Formulations and methods for lyophilization and lyophilates provided thereby

ABSTRACT

The present invention provides compositions, methods for lyophilizing compounds and making pharmaceutical compositions, and kits providing solutions and lyophilized formulations of compounds. The compositions, methods, and kits are particularly useful in pharmaceutical applications involving therapeutic agents that have low solubility at low pH and medium pH values. Certain embodiments provide methods for lyophilizing compounds in liquid solutions, which include the steps of: a) preparing aqueous solutions of a compound of interest in the absence of buffer; b) adjusting the pH to high values of pH in order to increase the solubility of the compound of interest; and c) freeze-drying the solution to provide a lyophilized solid composition. Aqueous solutions including buffer are also disclosed. Lyophilized formulations, including micronized and non-micronized powders, are provided.

BACKGROUND

Compounds that are weak acids are often poorly soluble at low (e.g.,less than about pH 4 or pH 5) and at medium pH values (e.g., pH valuesof about 6 or 7). Such compounds are often difficult to use aspharmaceuticals due to, for example, poor solubility in pharmaceuticallyacceptable solutions. Thus, despite possible theoretical therapeuticefficacy, some compounds are not useful in clinical practice, or not asuseful as they might be if methods and compositions were available toprovide such compounds in pharmaceutically acceptable forms. Inaddition, formulations of such compounds may be unstable, and may bedifficult to store for use, even if it were possible to preparepharmaceutically acceptable formulations of such compounds.

Sulphonylureas and other compounds that act on sulphonylurea receptors(SURs) arc useful in medical treatment of diabetes and other disorders.SURs may be of different types, including, for example, sulphonylureareceptor type 1 (SUR1) and sulphonylurea receptor type 2 (SUR2).Compounds that act at SURs include sulphonylureas (such asglibenclamide) and other compounds (e.g., repaglinide and taglizide).Sulphonylureas and other compounds active at SURs include glibenclamide(also known as glibenclamide), 4-trans-hydroxy-glibenclamide,3-cis-hydroxy-glibenclamide, tolbutamide, repaglinide, nateglinide,meglitinide, midaglizole, LY397364, LY389382, glyclazide, glimepirideand other drugs or metabolites of drugs which interact with SURs.

In addition, ion channels such as potassium channels and non-selectivechannels may be associated with SURs (e.g., a NC_(Ca-ATP) channel; see,for example, U.S. Pat. No. 7,285,574, hereby incorporated by referencein its entirety, or an ATP-sensitive potassium channel (K_(ATP),channel)). Compounds active towards ion channels associated with SURsare also useful in medical treatments. Some compounds that act onnon-selective channels that may be associated with SURs include, forexample, pinkolant, flufenamic acid, mefanamic acid, niflumic acid,rimonabant, and SKF 9635. In addition, other compounds may act on oraffect the action of SURs and/or ion channels associated with SURS,including, without limitation, for example, steroids and steroidderivatives and related compounds such as estrogen, estradiol, estrone,estriol, genistein, diethystilbestrol, coumestrol, zearalenone,non-steroidal estrogens, and phytoestrogens.

Glibenclamide Solubility

Glibenclamide solubility in various solutions has been reported, and istypically reported as being very poorly soluble in buffered aqueoussolutions. For example, the solubility of glibenclamide in bufferedaqueous solutions has been reported by Glomme et al. (Glomme A, Marz J,Dressman J B. Comparison of a miniaturized shake-flask solubility methodwith automated potentiometric acid/base titrations and calculatedsolubilities. J Pharm Sci. 2005 January;94(1):1-16). The bufferedaqueous solution was made with distilled water to form a potassiumchloride (220 mM) solution buffered with potassium phosphate (29 mM),and the pH adjusted to pH 5, 6, or 7 with sodium hydroxide. Thesesolutions had osmolarities of between about 280 to 310 milliOsmolar andhad buffer capacities of about 10±2 milliEquvialents/L/pH. Glomme et al.report that glibenclamide is only sparingly soluble in such solutions,with extremely low solubilities at pH 2, 3, 5, 6, and 7, and relativelygreater (although still very low) solubilities at pH 8, 9 and 11.8.These solubilities are shown in TABLE 1:

TABLE 1 Solubility of Glibenclamide at 37° C. (aqueous). pH Solubility(mg/mL) 2 0.00007 3 0.00006 5 0.0001 6 0.00062 7 0.00562 8 0.0512 90.0986 11.8 0.5316

It can be seen that glibenclamide in such aqueous solutions is poorlysoluble, that the solubility is less at acidic pH, and that thesolubility increases by an order of magnitude from pH 6 to pH 7, from pH7 to pH 8, and from pH 8 to pH 11.8.

Similarly, low glibenclamide solubilities in aqueous solutions werereported by Kaiser et al. (Kaiser D G, Forist, A A. A review ofGlibenclamide Metabolism in Man and Laboratory Animals. Physical andAnalytical Chemistry Research, The Upjohn Company; 1975), withsolubilities of below 1 mg/mL at all measured pH values from pH 4 to pH9. Glibenclamide was dissolved in Britton-Robinson buffer.(Britton-Robinson buffer is an aqueous buffer solution includingphosphoric acid, acetic acid and boric acid, with the pH adjusted withsodium hydroxide.) These solubilities are reported in TABLE 2.

TABLE 2 Solubility of Glibenclamide at 27° C. (aqueous). pH Solubility(mg/mL) 4 0.004 6 0.005 7 0.011 8 0.080 9 0.600

Rydberg et al. (Rydberg T, Jonsson A, Roder M, Melander A. Hypoglycemicactivity of glibenclamide (Glibenclamide) metabolites in humans.Diabetes Care. 1994 September;17(9):1026-30) also reported aglibenclamide solubility of 0.5 mg/mL in a 0.1 M, pH 10phosphate-buffered aqueous solution (300 mOsm/L).

The following formulation for intravenous glibenclamide (1 mg/mL) wasdeveloped for a Mayo study (Schrage W G, Dietz N M, Joyner M. T. Effectsof combined inhibition of ATP-sensitive potassium channels, nitricoxide, and prostaglandins on hyperemia during moderate exercise. J ApplPhysiol. 2006 May;l00(5):1506-12. Epub 2006 Feb. 9):

Ingredient Amount Glibenclamide 500 mg Sodium Chloride 0.9% 450 mL 0.1 NSodium Hydroxide 50 mL Above formula makes 500 mL Type of container 5 mLamber vial Amount in each 5 mL Shelf life Unknown

The formulation can be prepared by: i) mixing sodium hydroxide andsodium chloride in water; ii) dissolving glibenclamide in the mixture,with slight warming to help dissolve it; iii) filter the solutionthrough a 0.22 micron filter into sterile 5 mL amber vials; iv) stopper,cap and crimp. Sterility can be tested by using a Millipore system, andwhile working in the laminar flow hood: i) pass the test solutionthrough the filter and flush with sterile saline injection three times;ii) crimp the hoses and inject the culture media into the container;iii) record the product information on form #11.31, and staple to thecompounding formula; iv) perform a LAL test using a 1:20 dilution; v)quarantine for 14 days and check daily for presence or absence ofgrowth; vi) record all culture results on the culture report form andthe Microbial Culture Journal.

Betageri el al. (Betageri, G. V. et. al. Enhancement of dissolution ofglibenclamide by solid dispersion and lyophilization techniques, Int. J.Pharm. 126, 155-160 (1995)) evaluated increasing solubility ofglibenclamide first by addition of various polyethylene glycol (PEG) andthen via various PEG forms plus lyophilization. Betageri did notlyophilize glibenclamide on its own, and the procedures were performedat pH 7.4 in buffered solutions. Glibenclamide-PEG was found to be moresoluble than glibenclamide alone. It is to be noted that all theBetageri formulations involve one or more PEG, and that theconcentrations are very low.

Lyophilization

Lyophilization is a term used to describe methods and actions thatprovide dried materials, such as powders, from liquids containing solidsor dissolved materials by freeze-drying (freezing a liquid containingdissolved or suspended material, and drying while frozen by sublimation)to provide a dry solid containing the dissolved or suspended material insolid form. Typically, aqueous solutions are used in lyophilization,although mixed aqueous/solvent solutions, and other liquid solutions,may be used. For example, a biological material may be lyophilized froma solution or suspension in which it is mixed with protective agents.Such a solution or suspension may then be frozen, and subsequentlydehydrated by sublimation. Sublimation may optionally be followed byfurther drying steps.

Many materials and chemicals may be lyophilized. For example, dilutechemicals, including organic molecules such as drugs, hormones,proteins, nucleic acids (e.g., DNA and RNA), lipids, and carbohydratesor other molecules, may be lyophilized to provide a dried form of achemical or mixture of chemicals. Biological samples may also belyophilized. Typically, lyophilization methods include freeze-drying aliquid solution or suspension to provide a dry residue containing a highconcentration of the dissolved or suspended compounds. In some cases,the solid provided by lyophilization may be or include a salt.

Lyophilization processes provide solids, such as powders, dried films,or cakes. Small particles may be obtained, if desired, from suchpowders, films, or cakes by procedures such as grinding or flaking.

However, some methods of lyophilization may be improved.

In addition, some materials may be difficult to lyophilize. Somematerials, including some organic molecules useful in pharmaceuticalapplications and as medicaments, are difficult to dissolve or suspend ina solution, particularly in aqueous solutions of neutral or near-neutralpH, or in buffered aqueous solutions.

Thus, the need exists for improved methods of lyophilizing materialssuitable for a wider range of materials than is presently available, andfor particular desired materials and for desired types of materials.

SUMMARY OF THE INVENTION

Methods, compositions, and kits providing solutions and lyophilizedformulations of compounds of interest are taught herein. Compounds ofinterest are often compounds that are poorly soluble at low and mediumpH values, although more soluble at higher pH values.

The methods, compositions, and kits provided herein providepharmaceutically acceptable formulations, including solutions andlyophilized formulations, that solve the low solubility and lowstability problems associated with pharmaceutical formulations ofcompounds that are poorly soluble at low and medium pH values.

Compounds of interest may be, for example, sulphonylurea compounds, ionchannel-blocking compounds, steroid compounds, and other compoundshaving pharmaceutical activity. For example, methods, compositions andkits providing lyophilized formulations of sulphonylurea compounds aretaught herein. In a further example, methods, compositions and kitsproviding lyophilized formulations of ion-channel blocking compounds aretaught herein. In a still further example, methods, compositions andkits providing lyophilized formulations of sulphonylurea compoundstogether with steroid compounds are taught herein. In a yet furtherexample, methods, compositions and kits providing lyophilizedformulations of sulphonylurea compounds together with ion-channelblocking compounds and/or steroid compounds are taught herein. Inanother example, methods, compositions and kits providing lyophilizedformulations of sulphonylurea compounds and/or ion channel blockingcompounds, optionally along with a substantially pharmaceutically inertcompound are taught herein.

Methods for lyophilizing compounds from liquid solutions, and productscomprising lyophilized solids obtained from such lyophilized liquidsolutions, are provided herein. In certain embodiments, hydrophobicorganic molecules are dissolved in aqueous solutions and lyophilized toprovide solid compositions containing high concentrations of thehydrophobic organic molecules. In certain embodiments, these solidcompositions containing high concentrations of hydrophobic organicmolecules are stable and are suitable for storage, e.g., suitable forstorage for long periods of time. Such storage may be at ambientconditions, may be under controlled temperature, may be under controlledhumidity, or other condition or set of conditions; and may be stored ina sealed container (e.g., a bottle or jar with a removable lid, a tubes,a capsule, a caplet, a vial, or other container), and may be in a sealedcontainer under an inert gas (e.g., nitrogen, argon, helium, or otherinert gas), or other container with or without other element or compoundin the container.

Sulphonylureas and other compounds active at sulphonylurea receptors(SURs) include many chemicals that may be difficult to dissolve orsuspend in an aqueous solution. Sulphonylureas and other compoundsactive at SURs include glibenclamide (also known as glibenclamide),tolbutamide, repaglinide, nateglinide, meglitinide, midaglizole,LY397364, LY389382, glyclazide, glimepiride and other drugs ormetabolites of drugs which interact with SURs. Other compounds which mayalso exhibit similar problems with going into solution, and which may besuitable for use in the practice of embodiments of the invention,include compounds termed herein “ion channel-blocking” compounds, suchas, for example, pinkolant, flufenamic acid, mefanamic acid, niflumicacid, rimonabant, and SKF 9635. In addition, steroids and steroidderivatives and related compounds may also be lyophilized followingembodiments of methods of the invention; such steroids, steroidderivatives and related compounds include, without limitation, estrogen,estradiol, estrone, estriol, genistein, diethystilbestrol, coumestrol,zearalenone, non-steroidal estrogens, and phytoestrogens. In addition,mixed solutions, containing combinations of these compounds, or thesecompounds in combination with other compounds, may be lyophilized inembodiments of the methods and compositions having features of theinvention.

In addition, solutions and lyophilized formulations having features ofthe invention may include compounds which act to maintain, or aid inmaintaining, proper levels of blood glucose, or which act to raise, oraid in raising, blood levels of glucose, preferably to maintain or raiseglucose levels in the blood of a subject at or near normal physiologicallevels. Such compounds include, for example, glucose itself, othercarbohydrates, glucagon, and other compounds, and combinations thereof.

As disclosed herein, glucose thus may be included in solutions andlyophilized formulations having features of the invention as an elementhaving pharmaceutical activity, and may also be included in solutionsand lyophilized formulations having features of the invention forpurposes other than as an element having pharmaceutical activity. Thus,glucose, which has physiological activity, and may act as apharmaceutically active element of solutions and lyophilizedformulations having features of the invention, may also be included insolutions and lyophilized formulations having features of the inventionfor purposes other than its physiological effects. For example, glucosemay be included, and may be discussed herein as a “pharmaceuticallyinert” compound or a “substantially pharmaceutically inert” compound.Such reference to glucose recognizes, e.g., its osmotic, bulking, orother properties in addition to its pharmaceutical activities. It willbe understood that glucose is an element that may be included both whereactive ingredients are discussed herein, and where substantiallypharmaceutically inert ingredients arc discussed herein.

In addition, solutions and lyophilized formulations having features ofthe invention may include compounds which are substantiallypharmaceutically inert. As used herein, compounds that are“substantially pharmaceutically inert” include sugars such as glucose,fructose, mannose, galactose, mannitol, sorbitol, lactose, sucrose,trehalose, and other sugars, including mono-saccharides, di-saccharides,and other sugars; salts such as sodium chloride, potassium chloride, andother compounds which may be used in pharmaceutical solutions withlittle or no effect on the pharmacological activity of an activeingredient, but which may have effects on, for example, the osmolarityof the solution in which the substantially pharmaceutically inertcompounds are included. Note that the designation of glucose as a“substantially pharmaceutically inert” compound is for convenience ofreference only; glucose has physiological activity and may be includedfor pharmaceutical activity at the same time as, or in addition to, itsutility as a “substantially pharmaceutically inert” compound.

Thus, for example, in embodiments, solutions and methods suitable forthe practice of the invention may include solutions, lyophilizedformulations, and kits including solutions and/or lyophilizedformulations of glibenclamide, 4-trans-hydroxy-glibenclamide,3-cis-hydroxy-glibenclamide, tolbutamide, chlorpropamide, tolazamide,repaglinide, nateglinide, meglitinide, midaglizole, tolazamide,gliquidone, LY397364, LY389382, glyclazide, glimepiride, estrogen,estradiol, estrone, estriol, genistein, diethystilbestrol, coumestrol,zearalenone, non-steroidal estrogens, phytoestrogens, pinkolant,flufenamic acid, mefanamic acid, niflumic acid, rimonabant. SKF 9635,and combinations thereof. Furthermore, solutions and methods suitablefor the practice of the invention may include solutions, lyophilizedformulations, and kits including solutions and/or lyophilizedformulations of glibenclamide, 4-trans-hydroxy-glibenclamide,3-cis-hydroxy-glibenclamide, tolbutamide, chlorpropamide, tolazamide,repaglinide, nateglinide, meglitinide, midaglizole, tolazamide,gliquidone, LY397364, LY389382, glyclazide, glimepiride, estrogen,estradiol, estrone, estriol, genistein, diethystilbestrol, coumestrol,zearalenone, non-steroidal estrogens, phytoestrogens, pinkolant,flufenamic acid, mefanamic acid, niflumic acid, rimonabant, SKF 9635,and combinations thereof in which one or more substantiallypharmaceutically inert compound is also present in the solution,lyophilized formulation, or kit.

Methods for lyophilizing compounds in liquid solutions may include stepsof: a) preparing aqueous solutions of a compound of interest in theabsence of buffer or in the presence of a weak buffer (e.g., less thanabout 2 mM); b) adjusting the pH to high values of pH in order toincrease the solubility of the compound of interest; and c)freeze-drying the solution to provide a lyophilized solid composition.It will be understood that the term “compound of interest” as usedherein may be any one of the compounds named in the previous paragraph,and may include mixtures and combinations of more than one compound, andmay include mixtures and combinations including one or more of, forexample, the compounds named in the preceding paragraph. For example,methods for lyophilizing compounds in liquid solutions include preparingaqueous solutions of glibenclamide in the absence of buffer, adjustingthe pH to high values of pH in order to increase the solubility of theglibenclamide. In a further example, methods for lyophilizing compoundsin liquid solutions may include steps of preparing aqueous solutions ofglibenclamide and another compound such as, for example, pinkolant, inthe absence of buffer, adjusting the pH to high values of pH in order toincrease the solubility of these compounds. Adjusting the pH of thesolution to high pH values may be achieved using sodium hydroxide,potassium hydroxide, lithium hydroxide, calcium hydroxide, magnesiumhydroxide, or other agent suitable for increasing the pH of an aqueoussolution. Once the compound of interest (or mixture of compounds) isdissolved, the pH of the solution may optionally be lowered, e.g., byaddition of acid such as hydrochloric acid, sulfuric acid, acetic acid,or other acid. The resulting solution, prepared by any of the optionalmethods and having any of the optional compositions discussed herein,may be freeze-dried effective to provide a lyophilized solid. Thereconstituted liquid may have a lower pH than that of thepre-lyophilized solution due to the lyophilization process, with thecompound or compounds remaining in solution despite the lower pH.

Methods for lyophilizing compounds in liquid solutions may include stepsof: a) adding a compound of interest to water in the absence of buffer;b) adjusting the pH to high values of pH in order to increase thesolubility of the compound of interest; c) adding further amounts of acompound of interest; and d) freeze-drying the solution to provide alyophilized solid composition. Methods for lyophilizing compounds inliquid solutions may also include steps of: a) adding a compound ofinterest to water in the absence of buffer; b) adjusting the pH to highvalues of pH in order to increase the solubility of the compound ofinterest; c) adding further amounts of a compound of interest; d)repeating steps a), b), and c) one or more times as needed to achieve adesired, or to achieve a maximal, concentration of the compound ofinterest in the aqueous solution; and d) freeze-drying the solution toprovide a lyophilized solid composition.

Methods for lyophilizing compounds in liquid solutions may include stepsof: a) adding a compound of interest to water in the absence of bufferor in the presence of a weak buffer (less than about 2 mM); b) adjustingthe pH to high values of pH in order to increase the solubility of thecompound of interest; c) allowing the liquid to sit for a periodsufficient for the pH to reduce; (d) further adjusting the pH to highvalues of pH in order to increase the solubility of the compound ofinterest; and e) freeze-drying the solution to provide a lyophilizedsolid composition. Methods for lyophilizing compounds in liquidsolutions may also include steps of: a) adding a compound of interest towater in the absence of buffer; b) adjusting the pH to high values of pHin order to increase the solubility of the compound of interest; c)allowing the liquid to sit for a period sufficient for the pH to reduce;(d) further adjusting the pH to high values of pH in order to increasethe solubility of the compound of interest; e) repeating steps a), b),c) and d) one or more times as needed to achieve a desired, or toachieve a maximal, concentration of the compound of interest in theaqueous solution; and I) freeze-drying the solution to provide alyophilized solid composition.

The pH can be measured at any of the aforementioned steps, a single timeor multiple times. The pH can be adjusted at any of the aforementionedsteps, a single time or multiple times. A compound of interest may beadded to water, or to a water solution containing compound of interest,in the absence of buffer, a single time or multiple times. A compound ofinterest may be added to high-pH water, or a high-pH water solution, orto a high-pH water solution containing compound of interest, in theabsence of buffer, a single time or multiple times.

Thus, in embodiments of the methods, compositions and kits of theinvention, solutions and lyophilized formulations, and kits includingsuch solutions and lyophilized formulations, may include, for example,sulphonylureas and other compounds active at SURs such as glibenclamide,tolbutamide, repaglinide, nateglinide, meglitinide, midaglizole,LY397364, LY389382, glyclazide, glimepiride and other drugs ormetabolites of drugs which interact with SURs; may include ion channelblockers such as, for example, pinkolant, flufenamic acid, mefanamicacid, niflumic acid, rimonabant, and SKF 9635; may include estrogen,estradiol, estrone, estriol, genistein, diethystilbestrol, coumestrol,zearalenone, non-steroidal estrogens, phytoestrogens or other steroidcompound; and may also include one or more substantiallypharmaceutically inert compound such as, for example, glucose, fructose,mannose, galactose, mannitol, sorbitol, lactose, trehalose, sucrose, andother sugars, including mono-saccharides, di-saccharides, and othersugars, sodium chloride, potassium chloride, or other substantiallypharmaceutically inert compound.

In further particular embodiments, methods for lyophilizing compounds inliquid solutions include a) preparing, in the absence of buffer, aqueoussolutions of a compound of interest together with a substantiallypharmaceutically inert compound, b) adjusting the pH to high values ofpH in order to increase the solubility of the compound of interest, andc) freeze-drying the solution to provide a lyophilized solidcomposition. For example, methods for lyophilizing compounds in liquidsolutions include preparing aqueous solutions of glibenclamide andmannitol in the absence of buffer, adjusting the pH to high values of pHin order to increase the solubility of the glibenclamide and mannitolcompounds. In a further example, methods for lyophilizing compounds inliquid solutions include preparing aqueous solutions of glibenclamideand another compound in the absence of buffer, adjusting the pH to highvalues (e.g., pH of 8, 9, 10, or 11) in order to increase the solubilityof the compound of interest. Adjusting the pH of the solution to a highpH value may be achieved using sodium hydroxide, potassium hydroxide,lithium hydroxide, calcium hydroxide, magnesium hydroxide, or otheragent suitable for increasing the pH of an aqueous solution. Once thecompound of interest (or mixture of compounds) is dissolved, the pH ofthe solution may optionally be lowered, e.g., by addition of acid suchas hydrochloric acid, sulfuric acid, acetic acid, or other acid. Theresulting solution, containing a compound of interest together with asubstantially pharmaceutically inert compound, prepared by any of theoptional methods and having any of the optional compositions discussedherein, may be freeze-dried effective to provide a lyophilized solid.

In further embodiments, methods for lyophilizing compounds in liquidsolutions include a) preparing, in the presence of a buffer, aqueoussolutions of a compound of interest (which may comprise a mixture ofcompounds of interest), b) adjusting the pH to high values of pH inorder to increase the solubility of the compound of interest, and c)freeze-drying the solution to provide a lyophilized solid composition.Once the compound of interest is dissolved, the pH of the solution mayoptionally be lowered, e.g., by addition of acid. The resultingsolution, containing a compound of interest together with asubstantially pharmaceutically inert compound, prepared by any of theoptional methods and having any of the optional compositions discussedherein, may be freeze-dried effective to provide a lyophilized solid.

In further embodiments, methods for lyophilizing compounds in liquidsolutions include a) preparing, in the presence of a buffer, aqueoussolutions of a compound of interest (which may comprise a mixture ofcompounds of interest) together with a substantially pharmaceuticallyinert compound, b) adjusting the pH to high values of pH in order toincrease the solubility of the compound of interest, and c)freeze-drying the solution to provide a lyophilized solid composition.Once the compound of interest is dissolved, the pH of the solution mayoptionally be lowered, e.g., by addition of acid. The resultingsolution, containing a compound of interest together with asubstantially pharmaceutically inert compound, prepared by any of theoptional methods and having any of the optional compositions discussedherein, may be freeze-dried effective to provide a lyophilized solid.

Kits having features of the invention may include liquid solutions ofcompounds of interest, and/or liquid solutions of mixtures of compoundsof interest, and/or liquid solutions of compounds of interest togetherwith one or more substantially pharmaceutically inert compound, and mayinclude instructions for the use of such liquid solutions. For example,instructions for the use of such liquid solutions may includeinstructions for freeze-drying such solutions in order to obtain alyophilized formulation of the compound or compounds of interest.Alternatively, or in addition, kits having features of the invention mayinclude lyophilized formulations of compounds of interest, and/orlyophilized formulations of mixtures of compounds of interest, and/orlyophilized formulations of compounds of interest together with one ormore substantially pharmaceutically inert compound, and may includeinstructions for the use of such lyophilized formulations. For example,instructions for the use of such lyophilized formulations may includeinstructions for re-constituting such lyophilized formulations toprovide solutions, preferably sterile solutions, suitable for use inresearch and/or in pharmaceutical, medical, veterinary, or otherclinical application. Kits may include in separate vials of a pHadjuster or of pH adjusters which are able to reduce the pH of thereconstituted solution, where a “pH adjuster” is a compound able toalter the pH of a water solution (e.g., sodium hydroxide, potassiumhydroxide, hydrochloric acid, or other compounds effective to alter thepH of a water solution).

Solutions, formulations, lyophilates, and methods for making the same,as disclosed herein, are useful to provide, for example, materials thatmay be used as medicaments, and to prepare medicaments, for thetreatment of diseases, disorders, and conditions. Sulphonylureasolutions, formulations, lyophilates, and methods for making the same,may be used as medicaments, and to prepare medicaments for treating, forexample, stroke, brain trauma, spinal cord injury, ischemia (of thebrain, of the spinal cord, of the heart, and of other organs), and anyother disease or condition in which cells may express a SUR1-sensitivenon-selective channel such as the NC_(Ca-ATP) channel. Sulphonylureasolutions, formulations, lyophilates, and methods for making the same,may be used as medicaments, and to prepare medicaments for treating, forexample, diabetes, for treating diseases or conditions affecting K_(ATP)channels or which may be treated by modulating K_(ATP) channels, andother conditions. Accordingly, the materials disclosed herein provideimproved medicaments and treatments, and the methods disclosed hereinprovide improved methods for making medicaments and for treatingpatients.

Further embodiments relate to a lyophilized glibenclamide powdersubstantially free of buffer and including one or more substantiallypharmaceutically inert compounds. In certain instances, thesubstantially pharmaceutically inert compounds are independently a sugaror a salt. In certain instances, the substantially pharmaceuticallyinert compounds are independently glucose, fructose, mannose, galactose,mannitol, sorbitol, lactose, trehalose, sucrose, sodium chloride, orpotassium chloride. In certain instances, one of the substantiallypharmaceutically inert compounds is mannitol. In certain instances, thepowder includes only one substantially pharmaceutically inert compound,which is a sugar or a salt.

In certain instances, the substantially pharmaceutically inert compoundis mannitol. In certain instances, the amount of buffer is less than 1%w/w. In certain instances, the amount of buffer is less than 0.1% w/w.In certain instances, the amount of substantially pharmaceutically inertcompounds is less than 10% w/w. In certain instances, the amount ofsubstantially pharmaceutically inert compounds is less than 5% w/w. Incertain instances, the amount of substantially pharmaceutically inertcompounds is between 2% w/w and 6% w/w. In certain instances, theglibenclamide has a water solubility of at least about 0.01 mg/mL at 20°C. in an aqueous solution having a pH of 7. In certain instances, thesubstantially pharmaceutically inert compound is mannitol provided inthe aqueous solution from which the glibenclamide powder was lyophilizedin the amount of about 3 mg/100 mL (3%). In certain instances, thesubstantially pharmaceutically inert compound is glucose. In certaininstances, the substantially pharmaceutically inert compound is glucoseprovided in the aqueous solution from which the glibenclamide powder waslyophilized in the amount of about 3 mg/100 mL (3%). In certaininstances, the substantially pharmaceutically inert compound is a saltprovided in the aqueous solution from which the glibenclamide powder waslyophilized in the amount of less than about 10 mg/100 mL (10%). Incertain instances, the substantially pharmaceutically inert compound issodium chloride or potassium chloride.

In certain instances, lyophilized glibenclamide powder is substantiallyfree of agents enhancing the solubility of glibenclamide. Representativeagents that may enhance the solubility of glibenclamide includecyclodextrins and solubilizing polymers, such as polyethylene glycol. Incertain instances, lyophilized glibenclamide powder contains less than1% w/w of agents enhancing the solubility of glibenclamide. In certaininstances, lyophilized glibenclamide powder contains less than 0.1% w/wor less than 0.01% w/w of agents enhancing the solubility ofglibenclamide.

Further embodiments relate to a solid pharmaceutical compositionconsisting essentially of a lyophilized glibenclamide powder, an alkalimetal base, and optionally a substantially pharmaceutically inertbulking agent selected from the group consisting of a mono-saccharideand di-saccharide. In certain instances, the alkali metal base is sodiumhydroxide or potassium hydroxide, and the substantially pharmaceuticallyinert bulking agent is mannitol, glucose, fructose, mannose, galactose,sorbitol, lactose, trehalose, or sucrose. In certain instances, thesubstantially pharmaceutically inert bulking agent is mannitol. Incertain instances, the amount of substantially pharmaceutically inertbulking agent is between 2% w/w and 6% w/w. In certain instances, theamount of alkali metal base is less than 10% w/w. In certain instances,the amount of alkali metal base is less than 5% w/w.

Further embodiments relate to a pharmaceutical composition comprising alyophilized glibenclamide powder and another pharmaceutically activecompound, the composition being substantially free of buffer. In certaininstances, the other pharmaceutically active compound is4-trans-hydroxy-glibenclamide, 3-cis-hydroxy-glibenclamide, tolbutamide,chlorpropamide, tolazamide, repaglinide, nateglinide, meglitinide,midaglizole, tolazamide, gliquidone, LY397364, LY389382, glyclazide,glimepiride, estrogen, estradiol, estrone, estriol, genistein,diethystilbestrol, coumestrol, zearalenone, non-steroidal estrogens,phytoestrogens, pinkolant, flufenamic acid, mefanamic acid, niflumicacid, rimonabant, or SKF 9635. In certain instances, the otherpharmaceutically active compound is 4-trans-hydroxy-glibenclamide,3-cis-hydroxy-glibenclamide, tolbutamide, repaglinide, nateglinide,meglitinide, midaglizole, LY397364, LY389382, glyclazide, orglimepiride.

Further embodiments relate to a method for lyophilizing a compound,comprising: a) preparing an aqueous solution of a compound in theabsence of buffer, b) adjusting the pH to greater than about 8 in orderto increase the solubility of the compound, and c) freeze-drying thesolution to provide a lyophilized solid composition. In certaininstances, the pH is greater than about 9. In certain instances, the pHis greater than about pH 10. In certain instances, the concentration ofthe compound in solution is greater than about 0.5 mg/mL. In certaininstances, the concentration of the compound in solution is greater thanabout 1 mg/mL. In certain instances, the concentration of the compoundin solution is greater than about 2 mg/mL. In certain instances, theconcentration of the compound in solution is greater than about 4 mg/mL.In certain instances, the compound is glibenclamide,4-trans-hydroxy-glibenclamide, 3-cis-hydroxy-glibenclamide, tolbutamide,chlorpropamide, tolazamide, repaglinide, nateglinide, meglitinide,midaglizole, tolazamide, gliquidone, LY397364, LY389382, glyclazide,glimepiride, estrogen, estradiol, estrone, estriol, genistein,diethystilbestrol, coumestrol, zearalenone, non-steroidal estrogens,phytoestrogens, pinkolant, flufenamic acid, mefanamic acid, niflumicacid, rimonabant, or SKF 9635. In certain instances, the compound isglibenclamide.

Further embodiments relate to an aqueous pharmaceutical compositioncomprising lyophilized glibenclamide powder, water, an alkali metalsalt, and a substantially pharmaceutically inert bulking agent selectedfrom the group consisting of a mono-saccharide and di-saccharide, thecomposition containing less than 1% w/v of a buffering agent. In certaininstances, the alkali metal salt is sodium chloride or potassiumchloride, and the substantially pharmaceutically inert bulking agent ismannitol, glucose, fructose, mannose, galactose, sorbitol, lactose,trehalose, or sucrose. In certain instances, the substantiallypharmaceutically inert bulking agent is mannitol. In certain instances,the amount of substantially pharmaceutically inert bulking agent isbetween 2% w/v and 15% w/v. In certain instances, the amount ofsubstantially pharmaceutically inert bulking agent is between 2% w/v and6% w/v. In certain instances, the amount of alkali metal salt is lessthan 5% w/v. In certain instances, the pH of the composition is about 6to about 8. In certain instances, the pH of the composition is 6.5 to8.0. In certain instances, the osmolality of the composition is 200 mOsmto 400 mOsm. In certain instances, the osmolality of the composition is250 mOsm to 330 mOsm. In certain instances, the composition furthercomprises glucose in the amount of 2% w/v to 10% w/v. In certaininstances, the substantially pharmaceutically inert bulking agent isglucose.

In certain embodiments, the compositions described herein includeglucose or a related carbohydrate, glucagon, or a combination thereof.Glucose or a related carbohydrate, glucagon, or a combination thereofmay serve as excipients. In certain embodiments, the glucose or relatedcarbohydrate, glucagon, or combination thereof may be present in anamount sufficient to provide a therapeutic effect and/or therapeuticbenefit along with the sulphonylurea compound. For example,co-administration of a sulphonylurea with a therapeutically effectiveamount of glucose or a related carbohydrate, glucagon, or combinationthereof may be helpful and/or effective to maintain appropriate levelsof serum glucose in the blood of a patient to which the formulation isadministered. Appropriate levels of blood glucose are, for example,within the range of about 60 mg/dl (milligrams per deciliter) to about150 mg/dl (about 3.3 mM (millimoles per liter) to about 8 mM glucose).Thus, glucose or a related carbohydrate, glucagon, or combinationsthereof when further combined with a sulphonylurea compound may beadministered in combination to maintain the serum glucose within thisrange while providing the therapeutic benefits of the sulphonylureacompounds.

For example, as disclosed in U.S. Pat. No. 7.285,574, administration ofsulphonylurea compounds to a subject in need thereof may be helpfuland/or effective to reduce the risk o stroke and/or hypoxia/ischemia,reduce the amount of damage following stroke and/or hypoxia/ischemia(e.g., reduce intracranial pressure, reduce cell death, reduce strokesize, and/or reduce spinal cord injury, etc). A suitable amount ofglucose, related carbohydrate, glucagon, or combination thereof,comprises an amount that maintains a reasonable level of blood glucosein the patient, for example, the amount of glucose, relatedcarbohydrate, glucagon, or a combination thereof maintains a bloodglucose level of at least about 60 mg/dl, more preferably, is effectiveto maintain blood glucose levels within an acceptable range, such as,for example, between about 60 mg/dl and about 150 mg/dl. Thus, theamount of glucose, related carbohydrate, glucagon, or a combinationthereof is helpful and/or effective to prevent the subject from becominghypoglycemic.

In certain embodiments, formulations having features of the inventionmay include a sulphonylurea compound or compounds, in combination withan additional therapeutic agent, such as tissue plasminogen activator(tPA) or functionally related compound, aspirin, statins, diuretics,warfarin, Coumadin, mannitol, etc. Further embodiments may includeformulations including a) a sulphonylurea compound or compounds, b) athrombolytic agent, and c) glucose, a related carbohydrate, glucagon, ora combination thereof.

Further embodiments relate to a method of treating a patient sufferingfrom a disorder selected from the group consisting of stroke, neuronalcell swelling, traumatic brain injury, spinal cord injury, organischemia, acute coronary syndrome, myocardial infarction, sepsis, anddiabetes, comprising administering intravenously to a patient in needthereof an effective amount of an aqueous pharmaceutical compositiondescribed herein. In certain instances, the disorder is stroke. Incertain instances, the patient is a human. In certain other instances,the disorder is stroke, ischemia, hypoxia/ischemia, spinal cord injury,brain trauma, or other brain injury. In certain embodiments, thecomposition administered to the patient comprises a sulphonylureacompound and optionally glucose, carbohydrate related to glucose,glucagon, or a combination thereof. In further embodiments, suchformulations may comprise a) a sulphonylurea compound or compounds, b)glucose, a carbohydrate related to glucose, glucagon, or a combinationthereof, and c) another therapeutic agent. In certain instances, theglucose, carbohydrate related to glucose, glucagon, or a combinationthereof is present in a therapeutically effective amount.

Further embodiments relate to a vial of glibenclamide powder packagedwith a vial of diluent. Further embodiments relate to a vial ofglibenclamide powder packaged with a vial of diluent, where the diluentis selected from the group consisting of water; water and alcohol; waterand glucose; and water, PEG and alcohol. In certain instances, thediluent is water and glucose. In certain instances, the glucose ispresent in the amount of between about 2% and about 15% as measured byweight per volume (w/v). In certain instances, the diluent is a 5%dextrose solution. Other embodiments relate to a vial of glibenclamidepowder packaged with a vial of diluent, where the pH of the diluent hasa pH of about 7.4 or greater. Still other embodiments relate to a vialof glibenclamide powder packaged with a vial of diluent, where thediluent has a pH of about pH 7.4 or greater and is buffered. In certaininstances, the buffer concentration is between about 1 mM and about 100mM. In certain instances, the buffer concentration is less than about 15mM. In certain instances, the buffer concentration is between about 5 mMand about 10 mM.

Further embodiments relate to a vial of glibenclamide powder packagedwith a vial of diluent, where the diluent has a pH of about 7.4 orgreater and is buffered with a pharmaceutically acceptable buffer.Further embodiments relate to a vial of glibenclamide powder packagedwith a vial of diluent, where the diluent has a pH of about 7.4 orgreater and is buffered with a buffer selected from meglumine anddiethanolamine. Further embodiments relate to a vial of glibenclamidepowder packaged with a vial of diluent, where the diluent has a pH ofabout 7.4 and is buffered with a buffer selected from meglumine anddiethanolamine. Further embodiments relate to a vial of micronizedglibenclamide powder packaged with a vial of diluent, where the diluenthas a pH of about 7.4 or greater and is buffered with a buffer selectedfrom meglumine and diethanolamine. Further embodiments relate to a vialof micronized glibenclamide powder packaged with a vial of diluent,where the diluent has a pH of about 7.4 and is buffered with a bufferselected from meglumine and diethanolamine. Further embodiments relateto a vial containing lyophilized glibenclamide, a bulking agent, and apH adjuster. Further embodiments relate to a vial containing lyophilizedglibenclamide, a bulking agent, and a pH adjuster, wherein the pH isadjusted using NaOH. Further embodiments relate to a vial containinglyophilized glibenclamide, a bulking agent, and a pH adjuster, whereinthe concentration of glibenclamide is about 1 mg/mL. In certaininstances, the bulking agent is mannitol. In certain instances, the pHprior to lyophilization is about pH 11.4. In certain instances, thestarting material is micronized glibenclamide. In certain instances, thevial is packaged with a vial of diluent (buffered or not) with a pH of7.4-8.0 to add to the product above following reconstitution to reducepH.

Further embodiments relate to a kit comprising: a lyophilizedformulation of a compound as described herein; a diluent solution; andinstructions for the use of such liquid solutions. In certain instances,said diluent solution is selected from water; water and alcohol; waterand polyethylene glycol (PEG); water and glucose; and water, alcohol andPEG. In certain instances, the diluent solution is water and glucose. Incertain instances, the glucose is present in the amount of between about2% and about 15% as measured by weight per volume (w/v). In certaininstances, the diluent is a 5% dextrose solution. In certain instances,the diluent comprising alcohol, where alcohol is ethanol. In certaininstances, where the pH of the diluent is about 7.4 or greater. Incertain instances, where the diluent is buffered. In certain instances,where the buffer concentration is between about 1 mM and about 100 mM.In certain instances, where the buffer concentration is less than about15 mM. In certain instances, where the buffer concentration is betweenabout 5 mM and about 10 mM. In certain instances, where the diluent hasa pH of about 7.4 or greater and is buffered with a pharmaceuticallyacceptable buffer.

Further embodiments relate to a liquid formulation consistingessentially of glibenclamide and water, the formulation having a pH thatis sufficiently high so that a change in pH of about 1 does not causeglibenclamide to precipitate from the solution. In certain instances,the formulation contains less than about 0.01% w/v of a buffer. Incertain instances, the formulation contains less than about 0.01% w/v ofa surfactant. In certain instances, the formulation contains less thanabout 0.01% w/v of a cosolvent. In certain instances, the pH of theformulation is greater than about 9. In certain instances, the pH of theformulation is greater than about 10. Further embodiments relate to alyophilate composition produced by lyophilizing one of the formulationsdescribed above. Still further embodiments relate to a a liquidpharmaceutical composition produced by reconstituting a lyophilatecomposition described herein, wherein the pH of said liquidpharmaceutical composition is sufficiently high to dissolve at leastabout 98% by weight of the lyophilate composition. In certain instances,the liquid pharmaceutical composition has a pH in the range of about 6to about 8, comprises saline, and at least 98% w/v of glibenclamide isdissolved.

Further embodiments relate to a liquid formulation consistingessentially of glibenclamide and water, the formulation having a pH thatis sufficiently high so that a change in pH of about 1 does not causeglibenclamide to precipitate from the solution, the formulation furthercharacterized in that the solution is suitable for lyophilization toform a lyophilate that can be reconstituted and diluted with saline toform a solution having a pH in the range about 6 to about 8 and theglibenclamide remains dissolved in solution. Other embodiments relate toa lyophilized glibenclamide powder produced by lyophilizing a liquidsolution consisting essentially of glibenclamide and one or moresubstantially pharmaceutically inert compounds, the liquid solutionhaving a pH greater than 9. In certain instances, the liquid solutioncontains less than about 0.01% w/v of a buffer. In certain instances,the liquid solution has a pH greater than 10.

Further embodiments relate to a liquid formulation consistingessentially of an active therapeutic agent and one or more substantiallypharmaceutically inert compounds, the liquid formulation having a pHgreater than 8, and wherein the active therapeutic agent is a weak acid.In certain instances, the liquid formulation contains less than about0.01% w/v of a buffer. In certain instances, the liquid formulation hasa pH greater than 9. In certain instances, the active compound is anorganic compound having a molecular weight of less than 500 g/mol andcomprising a sulphonylurea group. In certain instances, the activecompound is glibenclamide. Other embodiments relate to a lyophilizedglibenclamide powder produced by lyophilizing the liquid formulationdescribed above.

Formulations, compositions, and contents of kits as disclosed herein aresuitable as formulations and compositions, and/or for use in preparingpharmaceutical formulations and compositions, for administration to apatient in need of treatment. For example, a patient in need oftreatment may be a patient in need of treatment with an effective amountof an aqueous pharmaceutical composition described herein. A patient inneed of treatment may be, for example, any patient for whom asulphonylurea compound may provide therapeutic benefit, including, forexample, a patient suffering from diabetes, ischemia, hemorrhage, orother disorder or condition susceptible of treatment with asulphonylurea compound. A patient in need of treatment may be, forexample, any patient for whom a combination of a sulphonylurca compoundtogether with an ion-channel blocking compound may provide therapeuticbenefit. A patient in need of treatment may be, for example, any patientfor whom a combination of a sulphonylurca compound together with anion-channel blocking compound and/or steroid compound may providetherapeutic benefit. In another example, a patient in need of treatmentmay be, for example, any patient for whom a combination of asulphonylurea compound and/or an ion channel blocking compound, alongwith a substantially pharmaceutically inert compound may providetherapeutic benefit.

A patient in need of treatment may be, for example, a patient sufferingfrom diabetes, or from hemorrhage, or other disorder or condition. Apatient in need of treatment may be, for example, a patient sufferingfrom ischemia of any organ, or organs, or system. Such a system may be,for example, the nervous system, including a portion of the nervoussystem, or the cardiovascular system, or a part of the cardiovascularsystem. Such an organ may be, for example, the brain, the heart, amuscle, or other organ. A patient in need of treatment may be anypatient who may benefit from administration of the formulations,compositions, and/or contents of the kits disclosed herein. Furtherexamples of a patient in need of treatment include patients sufferingfrom a disorder selected from the group consisting of stroke,hemorrhage, neuronal cell swelling, traumatic brain injury, spinal cordinjury, organ ischemia, acute coronary syndrome, myocardial infarction,sepsis, and diabetes.

Another aspect of the invention relates to methods of processing a claimunder a health insurance policy in general, the processing of aninsurance claim for the coverage of a medical treatment or drug therapyinvolves notification of the insurance company, or any other entity,that has issued the insurance policy against which the claim is beingfiled, that the medical treatment or drug therapy will be performed. Adetermination is then made as to whether the medical treatment or drugtherapy that will be performed is covered under the terms of the policy.If covered, the claim is then processed, which can include payment,reimbursement, or application against a deductible. Accordingly, certainembodiments relate to a method for processing a claim under a healthinsurance policy submitted by a claimant seeking reimbursement for costsassociated with treatment using a composition or kit described herein,the method comprising: a) reviewing said claim; b) determining whethersaid treatment is reimbursable under said insurance policy; and c)processing said claim to provide partial or complete reimbursement ofsaid costs.

For example, embodiments of the invention include methods for processingclaims for medical insurance and/or reimbursement for purchase orprescription of any of the formulations, compositions and kits disclosedherein. In specific embodiments, the methods employ a computer for saidprocessing of an insurance claim and/or for reimbursement for purchaseor for prescription of any of the formulations, compositions and kitsdisclosed herein. Further embodiments relate to methods for processingclaims for medical insurance and/or reimbursement for purchase orprescription of any of the formulations, compositions and kits disclosedherein for treating a patient in need of treatment.

As disclosed herein, a method for processing claims for medicalinsurance and/or reimbursement for purchase or prescription of any ofthe formulations, compositions and kits disclosed herein may include thesteps of:

-   i) receiving a claim for medical insurance and/or reimbursement for    purchase or prescription of a formulation, composition, or kit as    disclosed herein; and-   ii) providing reimbursement for the medical treatment, procedure,    and/or medicament.

In a further embodiment, a method for processing claims for medicalinsurance and/or reimbursement for purchase or prescription of any ofthe formulations, compositions and kits disclosed herein may include thesteps of:

-   i) receiving a claim for medical insurance and/or reimbursement for    purchase or prescription of a formulation, composition, or kit as    disclosed herein;-   ii) evaluating the claim; and-   iii) providing reimbursement for the medical treatment, procedure,    and/or medicament.

In embodiments of these methods for processing an insurance claim, anyone or more of the steps may involve the use of a computer; any one ormore of the steps may involve the use of electronic data transfer; anyone or more of the steps may involve the use of a telephone and/orfacsimile device; any one or more of the steps may involve the use ofmail and/or of a delivery service; and any one or more of the steps mayinvolve the use of electronic fund transfer devices and/or methods.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows glibenclamide concentration (shown in units of mg/mL on thevertical axis) plotted as a function of pH in an aqueous sucrosesolution (2% sucrose in water, where “%” means g/100 mL) and in wateralone.

FIG. 2 shows the amount of glibenclamide in solution (in mg/mL) atvarious pH values, plotted with glibenclamide amounts on the verticalaxis, and pH shown as increasing from left to right along the horizontalaxis.

FIG. 3 shows the amount of glibenclamide in solution (in mg/mL) insaline, sucrose, and buffered (Britton Robinson) water solutions atvarious pH values, plotted with glibenclamide amounts on the verticalaxis, and pH shown as increasing from left to right along the horizontalaxis.

FIG. 4 is a table listing characterization data obtained from a studyevaluating the stability of lyophilized glibenclamide as described inexample 7.

FIG. 5 is a table listing characterization data obtained from a studyevaluating the stability of lyophilized glibenclamide as described inexample 7.

FIG. 6 is a table listing characterization data obtained from a studyevaluating the stability of lyophilized glibenclamide as described inexample 7.

FIG. 7 is a table listing characterization data obtained from a studyevaluating the stability of lyophilized glibenclamide as described inexample 7.

FIG. 8 is a table listing characterization data obtained from a studyevaluating the stability of lyophilized glibenclamide as described inexample 7.

DETAILED DESCRIPTION

Methods for lyophilization, solutions suitable as starting materials forlyophilization, dry materials and formulations resulting fromlyophilization, including powders, cakes, films, and salts, and othermethods, materials, and formulations are provided herein.

An exemplary compound which may be formulated into a solution suitablefor lyophilization according to embodiments of the methods disclosedherein is glibenclamide. Glibenclamide(5-chloro-N-[2-[4-(cyclohexylcarbamoylsulfamoyl)phenyl]ethyl]-2-methoxy-benzamide; also known as glibenclamide) is asulphonylurea compound having a molecular weight of 494 g/mol, a pKa of6.8, a melting point of about 169-174° C., and has the followingchemical structure:

Glibenclamide has a water solubility of 4 mg/L and an ethanol solubilityof 5 mg/mL. As such, glibenclamide is about three orders of magnitudemore soluble in ethanol than in water. The terms “glibenclamide” and“glibenclamide” are intended to encompass both the free base compoundand pharmaceutically acceptable basic salts thereof. In certainembodiments, the “glibenclamide” and “glibenclamide” are presentsubstantially in the form of the free base compound.

It is desirable to provide aqueous solutions of glibenclamide and ofother drugs and compounds that are only sparingly soluble in aqueoussolutions. For example, it is desirable to provide aqueous solutions ofglibenclamide, 4-trans-hydroxy-glibenclamide,3-cis-hydroxy-glibenclamide, tolbutamide, chlorpropamide, tolazamide,repaglinide, nateglinide, meglitinide, midaglizole, tolazamide,gliquidone, LY397364, LY389382, glyclazide, glimepiride, estrogen,estradiol, estrone, estriol, genistein, diethystilbestrol, coumestrol,zearalenone, non-steroidal estrogens, phytoestrogens, pinkolant,flufenamic acid, mefanamic acid, niflumic acid, rimonabant, SKF 9635,and combinations thereof.

For example, glibenclamide solutions, formulations, and lyophilates maybe prepared, including solutions and formulation which may be watersolutions of glibenclamide, without sugars, salts, or buffers; may bewater solutions of glibenclamide also including a sugar (e.g., one ormore of glucose, fructose, mannose, galactose, mannitol, sorbitol,lactose, trehalose, sucrose, and other sugars, includingmono-saccharides, di-saccharides, and other sugars), may be watersolutions of glibenclamide also including a salt (e.g., sodium chlorideor potassium chloride), may be water solutions of glibenclamide alsoincluding a buffer (e.g., a Britton-Robinson buffer, a phosphate buffer,a “Tris” buffer (containing Tris(hydroxymethyl)aminomethane), a HEPESbuffer (containing N-(2-Hydroxyethyl)piperazine-N′-(2-ethanesulfonicacid), or other buffer). It will be understood that such solutions,formulations, and lyophilates made from such solutions and formulations,may include combinations of the above.

As discussed above. Rydberg et al. report formulations of glibeneclamidein phosphate-buffered solutions. However, attempts by the presentinventor to prepare formulations similar to those of Rydberg et al.revealed that phosphate-buffered glibenclamide lyophilizates did notreconstitute sufficiently. In addition, attempts by the present inventorto replicate the formulation of Schrage et al. (as reported in the Mayostudy) were unsuccessful, possibly because the glibenclamide would notdissolve sufficiently.

In contrast to previous pH-adjusted, lyophilized products that use pHstabilizers or buffers to keep the pH constant, one aspect of theinvention features a glibeneclamide formulation lacking a bufferingagent because it has been surprisingly discovered that such formulationsare better suited for lyophilization and subsequent reconstitution tomake a formulation suitable for intravenous administration. It has beenfound that a reduction in pH during lyophilization of glibeneclamideformulations described herein does not adversely affect the ability toreconstitute the end product. Moreover, the invention overcomes adisadvantage of certain previously described buffered solutions—thedisadvantage being that such buffered solution are often not suitablefor dilution following reconstitution to form solutions with suitable pHvalues for extended-length intravenous infusion (e.g., 1 day, 3 days, 5days). It is noted that very lightly buffered solutions (e.g., less thanabout 2 mM of buffering agent) are contemplated to be amenable to thepresent invention.

Accordingly, new methods and compositions providing aqueous solutions ofglibenclamide and of other drugs and compounds that are otherwise onlysparingly soluble in aqueous solutions are described herein. Exemplarymethods, solutions, formulations, lyophilates, and compositions aredescribed with particularity in the following examples.

The invention having been described generally will be described inreference to the various embodiments described below. The embodimentsdescribed below are presented for the purpose of further describingvarious aspects of the invention and should not be construed as limitingthe scope of the invention.

One aspect of the invention relates to a formulation suitable forlyophilization which contains glibenclamide, one or more pH adjustersand a bulking agent. In certain instances, the pH adjuster is NaOH andthe bulking agent is mannitol, sucrose, lactose or trehalose. In certaininstances, the concentration of glibenclamide is 1 mg/mL, and the pH is11.4. In certain instances, the pH adjusters are NaOH and HCl and thebulking agent is mannitol, sucrose, lactose or trehalose. In certaininstances, the concentration of glibenclamide is 0.2 mg/mL and the pH is9.4.

Another aspect of the invention relates to a vial containing thelyophilized contents o the above formulation. In certain instances, saidvial is packaged with a vial of diluent. In certain instances, thediluent has a pH of about pH 7.4 or greater and is buffered.

Another aspect of the invention relates to a method of preparing asolution suitable for lyophilization of a drug active at a sulphonylureareceptor (SUR), comprising the steps of: a) preparing a water solutionhaving a pH of about pH 8 or greater; b) adding a drug to the watersolution; and e) adjusting the pH of the resulting solution to have a pHof about 8 or greater; whereby a solution suitable for lyophilization ofa drug active at a sulphonylurea receptor (SUR) is obtained. In certaininstances, the pH is about 9 or greater.

Another aspect of the invention relates to a vial of Glibenclamidepowder packaged with a vial of diluent. Certain other aspects of theinvention relate to a vial of Glibenclamide powder packaged with a vialof diluent, where the diluent is selected from the group consisting ofwater; water and alcohol; and water, PEG and alcohol. Certain otheraspects of the invention relate to a vial of Glibenclamide powderpackaged with a vial of diluent, where the pH of the diluent has a pH ofabout pH 7.4 or greater. Still other aspects of the invention relate toa vial of Glibenclamide powder packaged with a vial of diluent, wherethe diluent has a pH of about pH 7.4 or greater and is buffered. Incertain instances, the buffer concentration is between about 1 mM andabout 100 mM. In certain instances, the buffer concentration is lessthan about 15 mM. In certain instances, the buffer concentration isbetween about 5 mM and about 10 mM.

Another aspect of the invention relates to a vial of glibenclamidepowder packaged with a vial of diluent, where the diluent has a pH ofabout pH 7.4 or greater and is buffered with a pharmaceuticallyacceptable buffer. Certain other aspects of the invention relate to avial of Glibenclamide powder packaged with a vial of diluent, where thediluent has a pH of about pH 7.4 or greater and is buffered with abuffer selected from meglumine and diethanolamine. Still other aspectsof the invention relate to a vial of Glibenclamide powder packaged witha vial of diluent, where the diluent has a pH of about pH 7.4 or greaterand is buffered with a buffer selected from meglumine anddiethanolamine.

Another aspect of the invention relates to a glibenclamide powdersubstantially free of buffer. Certain other aspects of the inventionrelate to a glibenclamide powder substantially free of buffer andincluding a substantially pharmaceutically inert compound. In certaininstances, the substantially pharmaceutically inert compound is selectedfrom a sugar and a salt. In certain instances, the substantiallypharmaceutically inert compound is selected from glucose, fructose,mannose, galactose, mannitol, sorbitol, lactose, trehalose, sucrose, andother sugars, including mono-saccharides, di-saccharides, and othersugars, sodium chloride, and potassium chloride. In certain instances,the substantially pharmaceutically inert compound is mannitol. Incertain instances, the substantially pharmaceutically inert compound ismannitol provided in the aqueous solution from which the glibenclamidepowder was lyophilized in the amount of about 3 mg/100 mL (3%). Incertain instances, the substantially pharmaceutically inert compound isglucose. In certain instances, the substantially pharmaceutically inertcompound is glucose provided in the aqueous solution from which theglibenclamide powder was lyophilized in the amount of about 3 mg/100 mL(3%). In certain instances, the substantially pharmaceutically inertcompound is a sugar provided in the aqueous solution from which theglibenclamide powder was lyophilized in the amount of less than about 10mg/100 mL (10%). In certain instances, the substantiallypharmaceutically inert compound is a sugar provided in the aqueoussolution from which the glibenclamide powder was lyophilized in theamount of less than about 5 mg/100 mL (5%). In certain instances, thesubstantially pharmaceutically inert compound is a salt. In certaininstances, the substantially pharmaceutically inert compound is a saltselected from sodium chloride and potassium chloride. In certaininstances, the substantially pharmaceutically inert compound is a saltprovided in the aqueous solution from which the glibenclamide powder waslyophilized in the amount of less than about 10 mg/100 mL (10%). Incertain instances, the substantially pharmaceutically inert compound isa salt provided in the aqueous solution from which the glibenclamidepowder was lyophilized in the amount of less than about 5 mg/100 mL(5%). In certain instances, the substantially pharmaceutically inertcompound is a salt provided in the aqueous solution from which theglibenclamide powder was lyophilized in the amount of less than or equalto about 2 mg/100 mL (2%).

Another aspect of the invention relates to a glibenclamide powder(micronized or non micronized) substantially free of buffer andincluding another pharmaceutically active compound. In certaininstances, the other pharmaceutically active compound is selected from4-trans-hydroxy-glibenclamide, 3-cis-hydroxy-glibenclamide, tolbutamide,chlorpropamide, tolazamide, repaglinide, nateglinide, meglitinide,midaglizole, tolazamide, gliquidone, LY397364, LY389382, glyclazide,glimepiride, estrogen, estradiol, estrone, estriol, genistein,diethystilbestrol, coumestrol, zearalenone, non-steroidal estrogens,phytoestrogens, pinkolant, flufenamic acid, mefanamic acid, niflumicacid, rimonabant, and SKF 9635. In certain instances, the otherpharmaceutically active compound is selected from4-trans-hydroxy-glibenclamide, 3-cis-hydroxy-glibenclamide, tolbutamide,repaglinide, nateglinide, meglitinide, midaglizole, LY397364, LY389382,glyclazide, and glimepiride. In certain instances, the otherpharmaceutically active compound is selected from pinkolant, flufenamicacid, mefanamic acid, niflumic acid, rimonabant, and SKF 9635. Incertain instances, the other pharmaceutically active compound isselected from estrogen, estradiol, estrone, estriol, genistein,diethystilbestrol, coumestrol, zearalenone, non-steroidal estrogens, andphytoestrogens.

Another aspect of the invention relates to a method for lyophilizing acompound, comprising: a) preparing an aqueous solution of a compound ofinterest in the absence of buffer, b) adjusting the pH to high values ofpH in order to increase the solubility of the compound, and c)freeze-drying the solution to provide a lyophilized solid composition.

In certain instances, a high value of pH comprises a pH value greaterthan about pH 7.4. In certain instances, a high value of pH comprises apH value greater than about pH 8. In certain instances, a high value ofpH comprises a pH value greater than about pH 8.5. In certain instances,a high value of pH comprises a pH value greater than about pH 9. Incertain instances, a high value of pH comprises a pH value greater thanabout pH 9.5. In certain instances, a high value of pH comprises a pHvalue greater than about pH 10. In certain instances, the concentrationof the compound in solutions having high values of pH is greater thanabout 0.3 mg/mL. In certain instances, the concentration of the compoundin solutions having high values of pH is greater than about 0.5 mg/mL.In certain instances, the concentration of the compound in solutionshaving high values of pH is greater than about 1 mg/mL. In certaininstances, the concentration of the compound in solutions having highvalues of pH is greater than about 2 mg/mL. In certain instances, theconcentration of the compound in solutions having high values of pH isgreater than about 3 mg/mL. In certain instances, the concentration ofthe compound in solutions having high values of pH is greater than about4 mg/mL. In certain instances, the concentration of the compound insolutions having high values of pH is greater than about 5 mg/mL. Incertain instances, the compound of interest is selected fromglibenclamide, 4-trans-hydroxy-glibenclamide,3-cis-hydroxy-glibenclamide, tolbutamide, chlorpropamide, tolazamide,repaglinide, nateglinide, meglitinide, midaglizole, tolazamide,gliquidone, LY397364, LY389382, glyclazide, glimepiride, estrogen,estradiol, estrone, estriol, genistein, diethystilbestrol, coumestrol,zearalenone, non-steroidal estrogens, phytoestrogens, pinkolant,flufenamic acid, mefanamic acid, niflumic acid, rimonabant, and SKF9635. In certain instances, the compound of interest comprises more thanone pharmaceutically active compound.

Another aspect of the invention relates to a method for lyophilizing acompound, comprising: a) adding a compound of interest to water in theabsence of buffer to provide an aqueous solution of the compound ofinterest; b) adjusting the pH to high values of pH; c) adding anadditional amount of the compound of interest to said aqueous solutionwater of the compound of interest; and d) freeze-drying the solution toprovide a lyophilized solid composition. Certain other aspects of theinvention relate to a method for lyophilizing a compound, comprising: a)adding a compound of interest to water in the absence of buffer toprovide an aqueous solution of the compound of interest; b) adjustingthe pH to high values of pH; c) adding an additional amount of thecompound of interest to said aqueous solution water of the compound ofinterest; d) repeating steps a), b), and c) one or more times as neededto achieve a desired, or to achieve a maximal, concentration of thecompound of interest in the aqueous solution; and e) freeze-drying thesolution to provide a lyophilized solid composition.

In certain instances, the compound of interest is selected fromglibenclamide, 4-trans-hydroxy-glibenclamide,3-cis-hydroxy-glibenclamide, tolbutamide, chlorpropamide, tolazamide,repaglinide, nateglinide, meglitinide, midaglizole, tolazamide,gliquidone, LY397364, LY389382, glyclazide, glimepiride, estrogen,estradiol, estrone, estriol, genistein, diethystilbestrol, coumestrol,zearalenone, non-steroidal estrogens, phytoestrogens, pinkolant,flufenamic acid, mefanamic acid, niflumic acid, rimonabant, and SKF9635. In certain instances, the compound of interest comprises more thanone pharmaceutically active compound. In certain instances, the aqueoussolution comprises a substantially pharmaceutically inert compound. Incertain instances, the substantially pharmaceutically inert compound isselected from a sugar and a salt. In certain instances, thesubstantially pharmaceutically inert compound is selected from glucose,fructose, mannose, galactose, mannitol, sorbitol, lactose, trehalose,sucrose, and other sugars, including mono-saccharides, di-saccharides,and other sugars, sodium chloride, and potassium chloride. In certaininstances, the substantially pharmaceutically inert compound in theaqueous solution has a concentration of about 10 mg/100 mL (10%). Incertain instances, the substantially pharmaceutically inert compound inthe aqueous solution has a concentration of about 5 mg/100 mL (5%). Incertain instances, the substantially pharmaceutically inert compound inthe aqueous solution has a concentration of about 3 mg/100 mL (3%). Incertain instances, the substantially pharmaceutically inert compound inthe aqueous solution has a concentration of about 1 mg/100 mL (1%).

Another aspect of the invention relates to a kit comprising: a liquidformulation of a compound of interest as described herein; andinstructions for the use of such liquid solutions. Certain other aspectsof the invention relate to a kit comprising: a lyophilized formulationof a compound of interest as described herein; a diluent solution; andinstructions for the use of such liquid solutions. In certain instances,said diluent solution is selected from water; water and alcohol; waterand polyethylene glycol (PEG); water, alcohol and PEG. In certaininstances, the diluent solution comprises alcohol, where alcohol isethanol. In certain instances, the pH of the diluent has a pH of aboutpH 7.4 or greater. In certain instances, the diluent has a pH of aboutpH 7.4 or greater and is buffered. In certain instances, the bufferconcentration is between about 1 mM and about 100 mM. In certaininstances, the buffer concentration is less than about 15 mM. In certaininstances, the buffer concentration is between about 5 mM and about 10mM. In certain instances, the diluent has a pH of about pH 1 7.4 orgreater and is buffered with a pharmaceutically acceptable buffer.

Another aspect of the invention relates to a vial of Glibenclamidepowder (micronized or non micronized) packaged with a vial of diluent,where the diluent has a pH of about pH 7.4 or greater and is bufferedwith a buffer selected from meglumine and diethanolamine. Certain otheraspects of the invention relate to a vial of Glibenclamide powder(micronized or non micronized) packaged with a vial of diluent, wherethe diluent has a pH of about pH 7.4 or greater and is buffered with abuffer selected from meglumine and diethanolamine.

Another aspect of the invention relates to a glibenclamide powder(micronized or non micronized) comprising a buffer. Certain otheraspects of the invention relate to a glibenclamide powder (micronized ornon micronized) comprising a buffer and including a substantiallypharmaceutically inert compound. In certain instances, the substantiallypharmaceutically inert compound is selected from a sugar and a salt. Incertain instances, the substantially pharmaceutically inert compound isselected from glucose, fructose, trehalose, sucrose, mannose, galactose,mannitol, sorbitol, sodium chloride, and potassium chloride.

Another aspect of the invention relates to a glibenclamide powder(micronized or non micronized) comprising a buffer and including anotherpharmaceutically active compound. In certain instances, the otherpharmaceutically active compound is selected from4-trans-hydroxy-glibenclamide, 3-cis-hydroxy-glibenclamide, tolbutamide,chlorpropamide, tolazamide, repaglinide, nateglinide, meglitinide,midaglizole, tolazamide, gliquidone, LY397364, LY389382, glyclazide,glimepiride, estrogen, estradiol, estrone, estriol, genistein,diethystilbestrol, coumestrol, zearalenone, non-steroidal estrogens,phytoestrogens, pinkolant, flufenamic acid, mefanamic acid, niflumicacid, rimonabant, and SKF 9635. In certain instances, the otherpharmaceutically active compound is selected from4-trans-hydroxy-glibenclamide, 3-cis-hydroxy-glibenclamide, tolbutamide,repaglinide, nateglinide, meglitinide, midaglizole, LY397364, LY389382,glyclazide, and glimepiride. In certain instances, the otherpharmaceutically active compound is selected from pinkolant, flufenamicacid, mefanamic acid, niflumic acid, rimonabant, and SKF 9635. Incertain instances, the other pharmaceutically active compound isselected from estrogen, estradiol, estrone, estriol, genistein,diethystilbestrol, coumestrol, zearalenone, non-steroidal estrogens, andphytoestrogens.

Another aspect of the invention relates to a vial containing lyophilizedglibenclamide, a bulking agent, and a pH adjuster. Certain other aspectsof the invention relate to a vial containing lyophilized glibenclamide,a bulking agent, and a pH adjuster, wherein the pH is adjusted usingNaOH. Still other aspects of the invention relate to a vial containinglyophilized glibenclamide, a bulking agent, and a pH adjuster, whereinthe concentration of glibenclamide is about 1 mg/mL. In certaininstances, the bulking agent is mannitol. In certain instances, the pHprior to lyophilization is about pH 11.4. In certain instances, thestarting material is micronized glibenclamide. In certain instances,packaged with a vial of diluent (buffered or not) with a pH of 7.4-8.0to add to the product above following reconstitution to reduce pH.

Definitions

To facilitate an understanding of the present invention, a number ofterms and phrases are defined below.

As used herein, the term “patient” refers to organisms to be treated bythe methods of the present invention. Such organisms preferably include,but are not limited to, mammals (e.g., murines, simians, equines,bovines, porcines, canines, felines, and the like), and most preferablyincludes humans.

As used herein, the term “effective amount” refers to the amount of acompound (e.g., a compound of the present invention) sufficient toeffect beneficial or desired results. An effective amount can beadministered in one or more administrations, applications or dosages andis not intended to be limited to a particular formulation oradministration route. As used herein, the term “treating” includes anyeffect, e.g., lessening, reducing, modulating, ameliorating oreliminating, that results in the improvement of the condition, disease,disorder, and the like, or ameliorating a symptom thereof.

As used herein, the term “pharmaceutical composition” refers to thecombination of an active agent with a carrier, inert or active, makingthe composition especially suitable for diagnostic or therapeutic use invivo or ex vivo.

As used herein, the term “pharmaceutically acceptable carrier” refers tostandard pharmaceutical carriers, such as saline solution, water,emulsions (e.g., such as an oil/water or water/oil emulsions), andvarious types of wetting agents. The compositions can, in certaininstances, include stabilizers and preservatives. For examples ofcarriers, stabilizers and adjuvants. (See e.g., Martin, Remington'sPharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, Pa. [1975]).

As used herein, the term “pharmaceutically acceptable basic salt” refersto any pharmaceutically acceptable basic salts of a compound of thepresent invention which, upon administration to a subject, is capable ofproviding a compound of this invention or an active metabolite orresidue thereof. As is known to those of skill in the art, “salts” ofthe compounds of the present invention may be derived from inorganic ororganic bases. Examples of bases include, but are not limited to, alkalimetals (e.g., sodium and potassium) hydroxides, alkaline earth metals(e.g., magnesium), hydroxides, ammonia, and compounds of formula NW₄ ⁺,wherein W is C₁₋₄ alkyl, and the like.

Examples

The invention now being generally described, will be more readilyunderstood by reference to the following examples, which are includedmerely for purposes of illustration of certain aspects and embodimentsof the present invention, and are not intended to limit the invention.

Example 1 Lyophilized Glibenclamide

TABLE 3 Low concentration High concentration formulation formulationConcentration of 0.2 mg/mL (0.1 mg/mL for NaCl 1.0 mg/mL glibenclamidebulking agent) (0.4 to 8 mg/mL) (0.05 to 0.4 mg/mL) pH prior to 9.4 11.4lyophilization (7+) (11+) Bulking agents Lactose Mannitol (NaCl,Sucrose, Mannitol, (Sucrose, Lactose, Trehalose. Trehalose) No NaCl)Bulking agent 2% 3% w/v % (1%+) (1%+) pH Begin compounding by raising pHBegin compounding by raising pH Adjustments to 11.3 using NaOH, add to11.3 using NaOH, add glibenclamide and excipients, glibenclamide andexcipients, adjusting up to pH 11.3 regularly. adjusting up to pH 11.3regularly. Then reduce to 9.4 with HCl No HCl. Buffer No buffer Nobuffer (or can be lightly buffered, e.g., (or can be lightly buffered,e.g., 2 mM) 2 mM) Reconstitute WFI (water for injection) WFI pH after8.4 to 9.4 10 to 11.4 reconstitution (can drop as low as 7 and stillreconstitute) Dilution for Dilute 1:1 with 0.9% Saline Dilute 1:10 with0.9% Saline bolus (1:1+) (1:1+) Dilution for Further dilute until pH <8.0, i.e., Further dilute until pH < 8.0, i.e., infusion physiologicallysuitable. physiologically suitable. Solvents None None (or PEG oralcohol) (or PEG or alcohol) Variations tried 0.25 mg/mL, pH 9.5 and pH10.5 0.5 mg/mL, pH 11.3 successfully 0.20 mg/mL, pH 9.5 and pH 10.5 0.1mg/mL, pH 9.5 and pH 10.5 Key: ranges or alternatives arc shown inparenthesis. “Water for injection” includes purified, sterilized,filtered, and other water that is suitable and safe for administrationto a patient.

Example 2A Solubility Studies Using Micronized Glibenclamide Sucrose

Micronized glibenclamide in 2% sucrose (as a proxy for glucose,fructose, mannose, galactose, mannitol, sorbitol, lactose, trehalose,and other sugars, including mono-saccharides, di-saccharides) in waterwas prepared, as recorded in the left-hand column of Table 4. Inaddition, micronized glibenclamide was prepared in water without abulking agent (see Table 4, right column). These were unbufferedsolutions, so that 1) pH was adjusted to the target value, then 2)glibenclamide was added (which lowered the pH) and then 3) pH wasre-adjusted up to the target. This was done iteratively until the pH wasstable.

TABLE 4 Solutions of Glibenclamide (GLB). Sucrose Water pH GLB mg/ml pHGLB mg/ml 7.45 0.051 7.36 0.019 7.50 0.099 7.55 0.096 8.24 0.310 8.650.799 8.66 1.069 8.73 0.087 9.16 2.608 9.13 0.205 9.53 3.060 9.57 3.9689.74 4.103 10.28 4.871 11.30 3.416 11.19 4.673 11.23 3.811 11.44 4.84111.99 1.904 12.27 2.513

Glibenclamide concentration (shown in units of mg/mL on the verticalaxis) is plotted in FIG. 1 as a function of pH in an aqueous sucrosesolution (2% sucrose in water, where “%” means g/100 mL) and in wateralone.

The results for lactose and mannitol and other sugars (e.g., glucose,fructose, mannose, galactose, trehalose, sorbitol, and other sugars,including mono-saccharides, di-saccharides) is contemplated to besimilar to those shown for sucrose.

Saline

Micronized glibenclamide in 2% saline and in 0.9% saline in water wasprepared and analyzed (see Table 5). These solutions were unbufferedsolutions, so the following method was used: 1) pH was adjusted to thetarget, then 2) glibenclamide was added (which lowered the pH) and then3) pH was re-adjusted up to the target. This was done iteratively untilpH was stable. “GLB” indicates glibenclamide.

TABLE 5 Solutions of Glibenclamide (GLB). Saline 2% Saline 0.9% pH GLBmg/ml pH GLB mg/ml 7.24 0.012 7.97 0.028 8.34 0.087 8.070 0.024 8.890.214 8.440 0.109 8.94 0.233 9.180 0.435 9.49 0.205 9.920 0.420 10.310.215 10.480 0.456 10.98 0.192 11.060 0.445 11.49 0.194 11.610 0.53812.10 0.216 12.070 0.476

The amount of glibenclamide (in mg/mL) was plotted on the vertical axisin FIG. 2 as a function of pH (along the horizontal axis) for solutionsof glibenclamide in saline solutions in and in water.

As can be seen in FIG. 2, the solubility of glibenclamide was less issaline solutions, with higher concentrations of NaCl further reducingthe solubility of glibenclamide. That is, the maximal amounts ofglibenclamide in the 0.9% saline solution were greater than the maximalamounts of glibenclamide in the 2% saline solution.

Buffered Solutions

Buffered water solutions (Britton Robinson buffers) were used todetermine the amounts of glibenclamide soluble in buffered solutionswithout added salt or added sugar. Experiments using buffered water plusglibenclamide resulted in the following concentrations by pH:

TABLE 6 Buffered (Britton Robinson) pH GLB mg/mL 5.3 0.01 6.200 0.017.100 0.01 8.000 0.02 8.3 0.02 9.000 0.14 9.700 0.85 10.100 2.35

When compared with the 2% sucrose and water-only versions, it appearsthat the solubility is slightly shifted in the buffered version, andthat higher solubility can be achieved using NaOH adjustment withoutbuffering:

Solution Containing 3% Mannitol, 1 mg/ml, Glibenclamide, and having a pHof 11.3

Glibenclamide—Trial Formulations (JC No.: R08-02682) and Lyophilizationand Reconstitution (JC.: R08-02683)

Experimental Details

Two bulk solutions of Glibenclamide in deionized water (Formula A, 1mg/mL; Formula B, 0.5 mg/mL) were prepared, both with 3% mannitol.Dilutions were prepared from each bulk solution. The bulk solutions wereassessed for appearance and pH immediately after preparation (0 hours),and after 5 (bulk solutions only) and 24 hours. Ten vials were filledfrom each of the two bulk solutions and subsequently lyophilized. Twovials from each batch were reconstituted with deionized water andassessed for appearance and pH. In the following, note that dilutionswere made using 0.9% saline solution.

The results are summarized below:

Results and Discussion

BN 838-097 (Glibenclamide 1.0 mg/mL in 3% mannitol, pH 11.4) and BN838-099 (Glibenclamide 0.01 mg/mL (1:99 Dilution of BN 838-097))

Time = Initial (0 hours) Experiment No. Dilution Appearance pH 838-097Undiluted Clear colourless solution 11.5 838-099 1:99 Clear colourlesssolution 6.5

Time = 5 hours Experiment No. Dilution Appearance pH 838-097 UndilutedClear colourless solution 11.4

Time = 24 hours Experiment No. Dilution Appearance pH 838-097 UndilutedClear colourless solution 11.4 838-099 1:99 Clear colourless solution6.7

Samples assessed at 0, 5 and 24 hours were clear colourless solutionswith no precipitate. The pH of the undiluted and diluted samples werestable over 24 hours.

BN 838-101 (Glibenclamide 0.5 mg/mL in 3% mannitol, pH 11.4), BN 838-103(Glibenclamide 0.01 mg/mL (1:49 Dilution of BN 838-101)), and BN 838-105(0.003 mg/mL (1:2 Dilution of BN 838-103))

The pH on dilution of BN 838-101 was noted to be above pH 8 (BN838-103), therefore a further 1:2 dilution was done of BN 838-103(therefore BN 838-105).

Time Zero

Time = Initial (0 hours) Experiment No. Dilution Appearance pH 838-101Undiluted Clear colourless solution 11.4 838-103 1:49 Clear colourlesssolution 9.4 838-105 1:2 (of BN 838-103) Clear, colourless solution 6.5

Time: 4 Hours

Time = 4 hours Experiment No. Dilution Appearance pH 838-101 UndilutedClear colourless solution 11.4

Time: 24 Hours

Time = 24 hours Experiment No. Dilution Appearance pH 838-101 UndilutedClear colourless solution 11.4 838-103 1:49 Clear colourless solution8.4 838-105 1:2 (of BN 838-103) Clear colourless solution 7.5

Samples assessed at 0, 4 and 24 hours were clear colourless solutionswith no precipitate. The pH of the undiluted sample was stable over 24hours. The pH of the diluted sample (BN 838-103 (1:49 Dilution of EN838-101)), showed a drop in pH by approximately 1 pH unit over 24 hours.

As described above, batch 838-103 was further diluted 1:2 due to the pHof batch 838-103 being above pH. The pH of this diluted sample increasedby approximately 1 pH unit over 24 hours, but remained below pH.

Lyophilization and Reconstitution

Experiment No. Dilution Appearance (n = 2) pH (n = 2) 838-097 UndilutedClear colourless solution 11.4, 11.4 838-101 Undiluted Clear colourlesssolution 11.4, 11.4

As described above, vials were filled from each of the bulk solutionsabove and lyophilized. At the end of the drying cycle, the vials wereunloaded from the dryer, and 2 vials from each batch reconstituted withdeionized water.

The freeze dried cakes dissolved very rapidly upon addition of deionizedwater to give clear colourless solutions with no precipitate. The pH wasmeasured and found to be 11.4 for both batches. A similar exercise wasundertaken at another laboratory, and the resultant pH followingreconstitution of the lyophilized material was 10.7. Therefore, theseresults indicate that the formulations can be successfully lyophilized.

Example 2B

EXEMPLARY FORMULATION: Glibenclamide (1 mg/mL), NaOH, and water; theformulation having a pH of 11.3.

TABLE 7 API (active pharmaceutical Micronized Glibenclamide ingredient)(Cambrex) API Concentration 1.0 mg/mL pH 11.4 ± 0.1 pH adjustment NaOHBulking Agent Mannitol Bulking Agent % 3% Water WFI (water forinjection) Specific Exclusions No HCl

Preparation

Lyophilized glibenclamide was produced by the following method:

-   1. Adjusted 95 mL WFI to pH 11.4±0.1 using 0.2M sodium hydroxide    solution, while stirring at medium speed.-   2. Add mannitol and glibenclamide with continued stirring. Adjust pH    to 11.4±0.1.-   3. During stirring, adjust pH of solution to 11.4 using 0.2M sodium    hydroxide solution every few minutes. Time taken for glibenclamide    to go into solution is approximately 30 minutes.-   4. Add WFI to bring volume to 100 mL.-   5. Adjust pH to 11.4 if required.-   6. The solution of step 5 is lyophilized. A white or off-white cake    is formed. The lyophilized material has a sufficiently large surface    area that there is no need to further grind it.    Note: No HCl is used. Glibenclamide causes some lowering of the pH;    no further reduction in pH is needed.

However, if desired, a further, optional, grinding step after step 6 maybe performed to grind the lyophilized cake into a micronized powder.Such a grinding step may be useful, for example, with large amounts oflyophilate, and may be omitted in any case, and particularly where thecake is small in size, within a vial, or for any other reason.

In experiments performed by the applicant, micronized glibenclamide wasobtained from Cambrex (supplied in micronized form; obtained fromCambrex Profarmaco Milano, Srl, Via Curiel, 34, 20067 Paullo (MI),Italy, a division of Cambrex Corporation of East Rutherford, N.J.07073). This micronized glibenclamide was then put into solution, andthat solution was then lyophilized according to the methods disclosedherein.

Unbuffered Solutions and Formulations

Unbuffered solutions and formulations were prepared—that is, thesolutions or formulations lacking a buffer. However, it is contemplatedthat solutions and formulations containing only low concentrations ofbuffer (e.g., less than about 2 mM) or only weak buffers or buffers withlow buffering capacity, so that the solution pH was not well-regulated,if regulated at all, by a buffer, are amenable to the present solutionsand formulations.

In certain embodiments of the methods disclosed herein, solution pH isto be continually increased during the manufacturing process as moreglibenclamide goes into solution.

Even though the pH is high following reconstitution, when thereconstituted solution is diluted in 0.9% Saline (e.g., diluted 50 timesto about 100 times, for example), the pH comes down to physiologicallyacceptable levels. Thus, such reconstituted solutions, upon dilution,are suitable for use, for example, in a 3-day infusion. This suitabilityis surprising due to the lack of buffering, which allows the pH to fallto acceptable levels upon dilution in a physiologically acceptablesolution, without further effort on the part of those diluting thesolution.

No HCl (Hydrochloric Acid)

Use of NaOH to increase pH, together with use of HCl to reduce pH canreduce the collapse temperature during lyophilization. Accordingly, itis preferable to use only NaOH for pH adjustment. Thus, in certainembodiments, HCl is not used: i.e., there is no use of HCl to adjust thepH of a solution or formulation for lyophilization.

No PEG (Polyethylene Glycol) or other Organic Solvent

Unlike other examples of lyophilized glibenclamide in the art, the useorganic solvents is not required in the methods and compositionsdisclosed herein. The absence of organic solvents is believed to be anadvantage over prior art formulations, and may simplify the FDAregulatory pathway (e.g., simplify the efforts needed to obtainregulatory approval for the use of solutions, formulations, lyophilates,etc., according to the methods disclosed herein, for use in treatingpatients and in producing medicaments for the treatment of diseases andconditions of patients) and so allow for rapid adoption of these methodsin clinical applications.

Relationship Between Starting pH, Starting Concentration, Total Dose,and Maximum Infusible Volume Per Day

For a particular starting concentration (e.g., 0.5 mg/mL), the startingpH that is high enough to maintain that concentration (e.g., pH 11.3)and total intended dose (e.g., 3 mg per day), the maximum infusiblevolume per day (e.g., 1,000 mL) has to be sufficiently high that whenthe drug is diluted in 0.9% Saline to get to the maximum infusiblevolume per day, the dilution ratio (994:6, i.e., 166:1) is higher thanthe minimum dilution ratio required to reach a pH of less than or equalto pH 8.5 or most preferentially pH 7.0 (in this case, the minimumdilution ratio is somewhere between 50-100 based on the experimentsdiscussed above).

Example 3

Exemplary Formulation: Glibenclamide (0.2 mg/mL), NaOH, HCl, and water;the formulation having a pH of 9.4.

This formulation is prepared as described for the 1 mg/mL glibenclamideformulation of Example 2B except that HCl is used as final step beforelyophilization to reduce the pH to 9.4.

Example 4

Exemplary Formulation: Glibenclamide (0.1 mg/mL), NaOH, HCl, and saline;the formulation having a pH of 9.4.

This formulation is prepared as described for the 1 mg/mL glibenclamideformulation of Example 2B except that HCl is used as final step beforelyophilization to reduce the pH to 9.4.

In addition, this formulation, and others of similar concentration, isimmediately isotonic upon reconstitution and contains no carbohydrates,which is believed to provide a clinical advantage in treating strokepatients, for example, as clinicians are often not comfortable givingcarbohydrates to stroke patients who are often hyperglycemic.

Freeze-Drying (Lyophilization)

Freeze drying (also known as lyophilization) is a process in which amaterial is first frozen, and then dried by sublimation (by reducing theair pressure around the frozen solid) and adding enough heat to causethe frozen water in the material to sublime directly from the solidphase to gas, leaving a dried material. Lyophilates often appear as dryflakes or other particles, which may then be further broken into smallerparticles to form, for example, a powder. A thorough description oflyophilization is found in the book Lyophilization—Introduction andBasic Principles by Thomas Jennings (published by CRC Press LLC, BocaRaton, Fla., USA (1999), ISI3N: 9781574910810 and ISBN-10: 574910817).

The freeze-drying process may be thought of as including three steps:Freezing, Primary Drying, and Secondary Drying.

The first step, freezing, is, as its name implies, simply the process offreezing the material. The material should be cooled to a temperaturebelow the eutectic point (the lowest temperature at which the solid andliquid phase of the material coexist) of the material to freeze it andto insure that sublimation rather than melting will occur withsubsequent heating of the frozen material under vacuum or low pressure.Since the eutectic point occurs at the lowest temperature where thesolid and liquid phase of the material can coexist, freezing thematerial at a temperature below this point ensures that sublimationrather than melting will occur in the following steps. Cooling ofamorphous (glassy) materials (which lack a eutectic point) should be tobelow the critical temperature of the material.

For example, a material may be frozen in a freeze-drying flask cooled byany suitable method (e.g., refrigeration, placement in a bath of dry iceand methanol, or placement in a liquid nitrogen bath).

In the initial drying step (the primary drying phase) the pressure islowered and enough heat is supplied for the frozen water in the materialto sublimate. Most of the water is removed in this initial drying phase.In this phase, pressure is controlled through the application of partialvacuum to speed sublimation.

Following the initial drying phase, further drying (the secondary dryingphase) is done by raising the temperature higher than the temperatureused in the primary drying phase. The secondary drying phase may removewater that has condensed or moved from an initial location during theprimary drying phase. Low pressure is typically used in this phase aswell.

Lyophilized products are often very stable, particularly if measures aretaken to prevent reabsorption of water. For example, lyophilization isuseful for providing pharmaceuticals that may be stored for many years.However, when needed, lyophilized materials products can be readilyrehydrated (reconstituted) as the process produces many microscopicpores in the material that aid reintroduction of water. Lyophilizedmaterial can be easily stored, shipped and later reconstituted to itsoriginal form for injection.

Micronizing

Reduction of the average particle size of a granulated or powdered solidmay be termed micronization, that is, for example, reducing the drug'sparticle size or micronizing the drug to have an average particle sizeof a few microns. It is often found that dosage forms which containmicronized drug particles exhibit enhanced solubility and consequentlyan increase in the bioavailability of the drugs.

Traditionally, dry materials were ground into fine powders (micronized)by hand by action of a mortar and pestle, in which the material wascrushed into finer and finer particles between the hard pestle and thehard mortar. Many mechanized micronization techniques (e.g., milling andgrinding) use friction to reduce particle size. A typical industrialmill is composed of a cylindrical metallic drum that contains grindingelements (e.g. steel spheres). As the drum rotates the grinding elementsinside the drum collide with the particles of the solid, and, whentrapped between two grinding elements, the particles are crushed toproduce smaller particles having smaller diameters. Alternatively,grinding wheels or other grinding elements may be used to micronizeparticles, such as powders or flakes, into smaller particles.

Methods like crushing and cutting are also used for reducing particlediameter, but produce rougher particles compared to milling and grinding(and are therefore the early stages of the micronization process).Crushing employs hammer-like tools to break the solid into smallerparticles by means of impact. Cutting uses sharp blades to cut the roughsolid pieces into smaller ones.

The micronization of solid materials, including proteins and drugs, toform solid particles suitable for microencapsulation (e.g., particleshaving an average particle size less than about less than 20 or lessthan about 10 μm) has been achieved using a variety of approachesincluding milling, as discussed above, and by spray-drying, sprayfreeze-drying, and supercritical anti-solvent (SAS) precipitationtechniques as well.

Various milling techniques are known. For example, in U.S. Pat. No.5,952,008 to Backstrom et al. jet milling is used to produce particlessmaller than 10 μm for inhalation administration. U.S. Pat. No.5,354,562 to Platz et al. discloses solid particle aerosol formulationsof polypeptide drugs made by lyophilizing solutions of the drugs whichcontain milling stabilizers that inhibit degradation of the drug duringsubsequent milling. The lyophilized drug is milled in fluid energy millsthat have been fitted with abrasion resistant materials. The resultingparticles are between 0.5 to 4 μm when milled at high pressure andbetween 4 μm to 15 μm when milled at low pressure. U.S. Pat. No.5,747,002 to Clark et al.

discloses jet milling of sodium chloride to produce particles with asize distribution smaller than 7 μm.

U.S. Pat. No. 5,817,343 to Burke discloses a method for formingpolymer/drug microparticles by forming a polymer solution/insoluble drugmixture; removing solvent from the mixture to form a hard matrixcontaining the drug particles in polymer; and micronizing the matrix byfragmenting (e.g., grinding or milling) the matrix below theglass-transition point of the polymer.

Sonication is another technique employed to micronize particles. Forexample, U.S. Pat. No. 4,384,975 to Fong et al. discloses thepreparation of microspheres by solvent removal using sodium oleate asthe emulsifier. Micronization of core material by milling or ultrasonicprobe sonication of solid drug particles in polymer solution isdisclosed. Tracy, Biotechnol. Prog, 14:108 15 (1998) discloses atomizinggrowth hormone in solution using an ultrasonic nozzle, freezing thedispersed droplets in a slurry of frozen ethanol, and then lyophilizingto remove the non-solvent and harden the droplets. The resulting hollowspheres are further micronized by ultrasonic probe treatment to fragmentthe spheres, which fragments are then encapsulated.

Example 5 Water Formulations for Preparing Lyophilized Tolbutamide

Tolbutamide is 24 times more soluble in water than glibenclamide (109mg/L vs. 4 mg/L) and so is more easily used in water solutions. However,about 100 times as much tolbutamide as glibenclamide is needed to havethe same clinical effect as glibenclamide. Thus, although in someembodiments, similar amounts of tolbutamide as glibenclamide may beincluded in the formulations and lyophilates having features of theinvention, or greater tolbutamide (or other drugs, as desired, includingfor example, repaglinide, nateglinide, meglitinide, midaglizole,LY397364, LY389382, glyclazide, glimepiride and other drugs ormetabolites of drugs which interact with SURs; may include ion channelblockers such as, for example, pinkolant, flufenamic acid, mefanamicacid, niflumic acid, rimonabant, and SKF 9635; may include estrogen,estradiol, estrone, estriol, genistein, diethystilbestrol, coumestrol,zearalenone, non-steroidal estrogens, phytoestrogens or other steroidcompound) are included as compared to the amounts of glibenclamidediscussed above.

In addition, formulations may include mixtures of drugs (e.g.,glibenclamide plus pinkolant; or glibenclamide plus flufenamic acid; orglibenclamide plus mefanamic acid; or glibenclamide plus niflumic acid;or glibenclamide plus rimonabant; or glibenclamide plus SKF 9635; orglibenclamide plus estrogen; or glibenclamide plus estradiol; orglibenclamide plus estrone; or glibenclamide plus estriol; orglibenclamide plus genistein; or glibenclamide plus diethystilbestrol;or glibenclamide plus coumestrol; or glibenclamide plus zearalenone; orglibenclamide plus a non-steroidal estrogen; or glibenclamide plus aphytoestrogen; or glibenclamide plus another drug or metabolite of adrug which interacts with SURs).

TABLE 8 Tolbutamide Formulations. Low concentration High concentrationformulation formulation Concentration of 0.2 mg/mL (0.1 mg/mL for NaCl1.0 mg/mL Tolbutamide bulking agent) (0.4 to 8 mg/mL) (0.05 to 0.4mg/mL) pH prior to expected to be 9-10 expected to be 11 or morelyophilization Bulking agents Lactose Mannitol (or NaCl, Sucrose,Mannitol, or (or Sucrose, Lactose, or Trehalose) Trehalose. No NaCl)Bulking agent 2% to 3% 3% to 4% w/v % pH Begin compounding by raising pHBegin compounding by raising pH Adjustments to pH 11 or above usingNaOH, to 11 or above using NaOH, add add tolbutamide and excipients,tolbutamide and excipients, adjusting up to pH 11 regularly. adjustingup to pH 11 regularly as Then reduce to 9-10 with HCl. necessary. No HCladded. Buffer No buffer No buffer (or the solution can be lightly (orthe solution can be lightly buffered, e.g., 2 mM) buffered, e.g., 2 mM)Reconstitute WFI (water for injection) WFI pH after expected to be 8 to9 expected to be 10 to 12 reconstitution (can reduce pH to 7 and stillreconstitute) Dilution for Dilute 1:1 or greater with 0.9% Dilute 1:10or greater with 0.9% bolus Saline Saline Dilution for Further diluteuntil pH < 8.0, i.e., Further dilute until pH < 8.0, i.e., infusionphysiologically suitable. physiologically suitable. Solvents None None(or an alcohol or polyethylene (or an alcohol or polyethylene glycol)glycol) Key: ranges or alternatives are shown in parenthesis.

Similar to the glibenclamide solutions, formulations, and lyophilatesdiscussed above, tolbutamide solutions, formulations, and lyophilatesmay be prepared. Such tolbutamide solutions and formulations may bewater solutions of tolbutamide, without sugars, salts, or buffers; maybe water solutions of tolbutamide also including a sugar (e.g., one ormore of glucose, fructose, mannose, galactose, mannitol, sorbitol,lactose, trehalose, sucrose, and other sugars, includingmono-saccharides, di-saccharides, and other sugars), may be watersolutions of tolbutamide also including a salt (e.g., sodium chloride orpotassium chloride), may be water solutions of tolbutamide alsoincluding a buffer (e.g., a Britton-Robinson buffer, a phosphate buffer,a “Tris” buffer (containing Tris(hydroxymethyl)aminomethane), a HEPESbuffer (containing N-(2-Hydroxyethyl)piperazine-N′-(2-ethanesulfonicacid), or other buffer). It will be understood that such solutions,formulations, and lyophilates made from such solutions and formulations,may include combinations of the above.

Example 6 Water Formulations for Preparing Lyophilized Repaglinide

Repaglinide is another pharmaceutically active ingredient that acts onSURs and is suitable for the practice of the invention. Repaglinide hasa water solubility of 0.6 mg/mL at pH 9, which is a bit lower than thesolubility of glibenclamide in water at this pH. Accordingly,repaglinide water formulations are contemplated to be similar toglibenclamide water formulations.

TABLE 9 Repaglinide Formulations. Low concentration High concentrationformulation formulation Concentration 0.2 mg/mL (0.1 mg/mL for NaCl 1.0mg/mL of repaglinide bulking agent) (or 0.4 to 8 mg/mL) (or 0.05 to 0.4mg/mL) pH prior to expected to be 9-10 expected to be 11 or morelyophilization Bulking agents Lactose Mannitol (or NaCl, Sucrose,Mannitol, or (or Sucrose, Lactose, or Trehalose) Trehalose. No NaCl)Bulking agent 2% to 3% 3% to 4% % pH Begin compounding by raising pHBegin compounding by raising pH Adjustments to pH 11 or above usingNaOH, to 11 or above using NaOH, add repaglinide and excipients, addrepaglinide and excipients, adjusting up to pH 11 regularly. adjustingup to pH 11 regularly as Then reduce to 9-10 with HCl necessary. No HCladded. Buffer No buffer No buffer (can be lightly buffered e.g. (can belightly buffered e.g. 2 mM) 2 mM) Reconstitute WFI (water for injection)WFI pH after expected to be 8 to 9 expected to be 10 to 12reconstitution (can drop as low as 7 and still reconstitute) Dilutionfor Dilute 1:1 or greater with 0.9% Dilute 1:10 or greater with 0.9%bolus Saline Saline Dilution for Further dilute until pH < 8.0 i.e.Further dilute until pH < 8.0 i.e. infusion physiologically suitable.physiologically suitable. Solvents None None (or PEG or alcohol) (or PEGor alcohol) Key: ranges or alternatives are shown in parenthesis.

Similarly to the glibenclamide and tolbutamide solutions, formulations,and lyophilates discussed above, repaglinide solutions, formulations,and lyophilates may be prepared. Such repaglinide solutions andformulations may be water solutions of repaglinide, without sugars,salts, or buffers; may be water solutions of repaglinide also includinga sugar (e.g., one or more of glucose, fructose, mannose, galactose,mannitol, sorbitol, lactose, trehalose, sucrose, and other sugars,including mono-saccharides, di-saccharides, and other sugars), may bewater solutions of repaglinide also including a salt (e.g., sodiumchloride or potassium chloride), may be water solutions of repaglinidealso including a buffer (e.g., a Britton-Robinson buffer, a phosphatebuffer, a “Tris” buffer (containing Tris(hydroxymethyl)aminomethane), aHEPES buffer (containingN-(2-Hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid), or otherbuffer). It will be understood that such solutions, formulations, andlyophilates made from such solutions and formulations, may includecombinations of the above.

Similarly, solutions and formulations, and lyophilates made from suchsolutions and formulations, may be made from other drugs andpharmaceutically active compounds and ingredients. Thus, for example,similar solutions and formulations, and lyophilates made from suchsolutions and formulations, may be made from, e.g., nateglinide,meglitinide, midaglizole, LY397364, LY389382, glyclazide, glimepirideand other drugs or metabolites of drugs which interact with SURs; ionchannel blockers such as, for example, pinkolant, flufenamic acid,mefanamic acid, niflumic acid, rimonabant, and SKF 9635; estrogen,estradiol, estrone, estriol, genistein, diethystilbestrol, coumestrol,zearalenone, non-steroidal estrogens, phytoestrogens or other steroidcompound; or other pharmaceutically active compound.

Kits may be prepared including solutions, formulations, and lyophilateshaving features of the invention. For example, a kit may include aliquid formulation of a compound of interest (e.g., as discussed above,including, for example, glibenclamide, tolbutamide, repaglinide,nateglinide, meglitinide, midaglizole, LY397364, LY389382, glyclazide,glimepiride and other drugs or metabolites of drugs which interact withSURs; ion channel blockers such as, for example, pinkolant, flufenamicacid, mefanamic acid, niflumic acid, rimonabant, and SKF 9635; estrogen,estradiol, estrone, estriol, genistein, diethystilbestrol, coumestrol,zearalenone, non-steroidal estrogens, phytoestrogens or other steroidcompound; or other pharmaceutically active compound; and instructionsfor the use of such liquid solutions. For example, the instructions mayinclude methods and description as described above, including methodsfor providing water solutions containing an active pharmaceuticalingredient. In embodiments, the instructions may simply describe how onemay add appropriate amounts of water to a dry lyophilate to provide awater solution. In embodiments, the instructions may further describehow one may measure the pH of such a solution, and may describe how onemay adjust the pH of such a solution as desired or as appropriate, asdescribed above. In embodiments, the instructions may describe how onemay add appropriate further ingredients, including buffers, salts,excipients, extenders, or other ingredients, as described above, to sucha solution. In embodiments, the instructions may describe how one mayadd such a solution to a suitable solution for injection, either asbolus or for infusion, as described above.

In further embodiments, a kit may include a lyophilized formulation of acompound of interest (e.g., as discussed above); a diluent solution; andinstructions for the use of such liquid solutions. For example, such adiluent solution may be selected from water; water and alcohol (e.g.,ethanol); water and polyethylene glycol (PEG); water, alcohol and PEG.In embodiments, the pH of the diluent may be a pH of about pH 7.4 orgreater.

In further embodiments, the pH of the diluent is about pH 7.4 or greaterand the diluent is buffered; the buffer may be a pharmaceuticallyacceptable buffer. In yet further embodiments, the diluent of the kit isbuffered, and the buffer concentration is between about 1 mM and about100 mM. In embodiments, the buffer concentration may be less than about15 mM; and in embodiments, the buffer concentration is between about 5mM and about 10 mM.

Example 7 Stability of Lyophilized Glibenclamide

A study was carried out to assess the stability of lyophilizedglibenclamide to extended storage at various temperatures and relativehumidity. The data obtained from this study indicate that lyophilizedglibenclamide has good stability over at least 3 months at thetemperatures and relative humidity conditions tested. Experimentalprocedures and results from the study are described in detail below.

Experimental Procedures: Vials containing lyophilized glibenclamidepowder were used in this study. The lyophilized glibenclamide powder wasobtained by lyophilization of an aqueous mixture containing 4.7-5.0 mgglibenclamide (The experiment started with 6 mg of glibenclamide, butsome glibenclamide remained on the filter following filtration of theaqueous mixture. As a result, the filtrate used for lyophilizationcontained 4.7-5.0 mg of glibenclamide.), 180 mg mannitol, and sodiumhydroxide as needed to adjust the pH to11.3 prior to lyophilization.Lyophilization of this aqueous mixture removed 6 mL of water per vial.

The lyophilized glibenclamide powder was evaluated at the following timepoints for appearance, reconstitution time, p1-1 after reconstitution,moisture content, amount of glibenclamide as analyzed by HPLC, and theamount of substances related to glibenclamide:

Time=initial at 2-8° C.

Time=6 weeks at 2-8° C.

Time=6 weeks at 25° C. and 60% RH

Time 6 weeks at 40° C. and 75% RH

Time=6 weeks and 13 days at 25° C. and 60% RH

Time=6 weeks and 13 days at 40° C. and 75% RH

Time=3 months at 2-8° C.

Time=3 months at 25° C. and 60% RH

Time=3 months at 40° C. and 75% RH

The analyses were carried out as follows:

-   Appearance and Particulates: Lyophilized vials were inspected    visually (before and after reconstitution with 6 mL of water for    injection).-   Reconstitution Time: Reconstitution time of duplicate samples from 2    separate vials were measured after adding 6 mL of water for    injection.-   pH: The pH of duplicate samples from two separate vials used for the    reconstitution were measured.-   Assay for Amount of Glibenclamide and Assay for Substances Related    to Glibenclamide:

The assay for glibenclamide in injection samples was determined by anisocratic HPLC method. A Zorbax XDB-C18, 5.0vun, 150 mm×4.6 mm columnwas used, operated at 50° C., eluting with an acetonitrile/water/fonnicacid eluent. Methanol was used as diluent. The glibenclamide content wasassayed by comparison with similarly chromatographed referencesolutions. Related substances were evaluated as area% with reference tothe Glibenclamide peak at 230 nm.

-   Moisture Content Analyzed by Coulometric Karl Fischer: The moisture    content was evaluated by dispersing the lyophilized cake in benzyl    alcohol and analyzing this solution by coulometric Karl Fischer    titration. Results are reported as mg/vial.

Results:

Appearance and Particulates: At the 3 month time point, the appearancebefore and after reconstitution of the lyophilized glibenclamide powdershowed no change from the initial time point. Before reconstitution awhite to off-white cake, approximately 12-14 mm in depth was observed,with some small cracks on the surface. After reconstitution a clearcolourless solution free from visible particles and fibres was observed.The appearance 24 hours after reconstitution contained several smallfibres across all time points and conditions, but these fibres weredetermined to have entered during determination of the pH.

Reconstitution Time: There was no significant variation in thereconstitution time across all time points and conditions.

pH: There was no significant variation in pH across all time points andconditions, and all results were within the pH range of 10.4-11.4.

Assay for Amount of Glibenclamide and Assay for Substances Related toGlibenclamide: Each vial was reconstituted with 6 mL of methanol to givean expected concentration of 0.78-0.83 mg/mL glibenclamide (It wasdetermined that each vial of lyophilized glibenclamide powder contained4.7-5.0 mg of glibenclamide.). Results from the assay of glibenclamideupon storage were within the range of 0.78-0.83 mg/mL, except forsamples stored at 40° C. and 75% RH at the 6 week and 3 month timepoints, and one sample stored 3 months at 2-8° C. The concentration ofglibenclamide in the samples upon storage at 40° C. and 75% RH were both0.77 mg/mL—the reduction in glibenclamide content is likely due toelevated temperature and relative humidity compared to other samples.Samples stored for 3 months at a temperature of 2-8° C. had aglibenclamide concentration of 0.844 mg/mL and 0.762 mg/mL.

Three substances presumably related to glibenclamide were identifiedduring the stability study. The first substance had a HPLC relativeretention time of 0.19 and produced a quantifiable peak. The secondsubstance had a 111³1,C relative retention time of 0.33 and was presentat trace levels for the 6 week time point only. The third substance hada HPLC relative retention time of 0.41 and was present at trace levels.Notably, for samples stored at 2-8° C. and at 25° C. with 60% RH, therewere no significant changes in concentration of related substances whencompared to the initial time point. Samples stored at 40° C. and 75% RHhad slightly increased levels of the first substance (i.e., thesubstance having a HPLC retention time at 0.19 minutes; 0.19% area inthe sample stored for 3 months, as compared to 0.12% area at the initialtime point).

Moisture Content Determined by Coulometric Karl Fischer Titration: Therewas no significant variation in the moisture content across all timepoints and conditions. All mean results are within the range of 1.5-2.0mg/vial.

Data from this study are presented in FIGS. 4-8.

INCORPORATION BY REFERENCE

The entire disclosure of each of the patent documents and scientificarticles referred to herein is incorporated by reference for allpurposes.

Equivalents

The invention may be embodied in other specific forms without departingfrom the spirit or essential characteristics thereof. The foregoingembodiments are therefore to be considered in all respects illustrativerather than limiting the invention described herein. Scope of theinvention is thus indicated by the appended claims rather than by theforegoing description, and all changes that come within the meaning andrange of equivalency of the claims are intended to be embraced therein.

1-103. (canceled)
 104. A solid pharmaceutical composition comprising alyophilized glibenclamide powder substantially free of buffer, includingone or more substantially pharmaceutically inert compounds and one ormore alkali bases, and being substantially free of other agents thatenhance the solubility of glibenclamide.
 105. The solid pharmaceuticalcomposition of claim 104, wherein the one or more substantiallypharmaceutically inert compounds are independently selected from a sugarand a salt.
 106. The solid pharmaceutical composition of claim 104,wherein the substantially pharmaceutically inert compounds areindependently selected from glucose, fructose, mannose, galactose,mannitol, sorbitol, lactose, trehalose, sucrose, sodium chloride, andpotassium chloride.
 107. The solid pharmaceutical composition of claim104, wherein the alkali metal base is sodium hydroxide or potassiumhydroxide.
 108. The solid pharmaceutical composition of claim 104,wherein one of the substantially pharmaceutically inert compounds ismannitol and the alkali base is sodium hydroxide.
 109. The solidpharmaceutical composition of claim 104, wherein the amount of buffer isless than 1% w/w.
 110. The solid pharmaceutical composition of claim104, wherein the amount of substantially, pharmaceutically inertcompounds is less than 10% w/w.
 111. A method for lyophilizing acompound of interest that binds to a sulphonylurea receptor or thatbinds to an ion channel associated with a sulphonylurea receptor toprovide a lyophilized solid compound, comprising: a) preparing anaqueous solution of a said compound in the absence of buffer, (b) addinga substantially pharmaceutically inert compound as a bulking agent c)adjusting the pH to greater than about pH 8 effective to increase thesolubility of the compound of interest, and d) freeze-drying thesolution to provide a lyophilized solid composition.
 112. The method ofclaim 111, wherein the pH is adjusted to greater than about pH
 10. 113.The method of claim 111, wherein the concentration of the compound ofinterest in solution is greater than about 0.5 mg/mL.
 114. The method ofclaim 111, wherein the substantially pharmaceutically inert compound ismannitol in the amount of less than about 10 mg/100 mL (10%).
 115. Themethod of claim 111, wherein the substantially pharmaceutically inertcompound is mannitol in the amount of about 3 mg/100 mL (3%).
 116. Anaqueous pharmaceutical composition comprising lyophilized glibenclamidepowder, wherein said lyophilized glibenclamide powder consistsessentially of glibenclamide, mannitol, and sodium hydroxide, saidlyophilized glibenclamide powder being mixed with water and/or saline.117. A method of treating a patient suffering from a disorder selectedfrom the group consisting of stroke, neuronal cell swelling, traumaticbrain injury, spinal cord injury, ischemia, hypoxia/ischemia, organischemia, acute coronary syndrome, myocardial infarction, sepsis, anddiabetes, comprising administering intravenously to a patient in needthereof an effective amount of an aqueous pharmaceutical composition ofclaim
 116. 118. The method of claim 117, wherein the disorder isselected from the group consisting of stroke, neuronal cell swelling,traumatic brain injury, spinal cord injury, ischemia, andhypoxia/ischemia.
 119. A liquid formulation consisting essentially ofglibenclamide, a bulking agent, an alkali base and water, theformulation having a pH that is sufficiently high so that a change in pHof about 1 does not cause glibenclamide to precipitate from thesolution, wherein said sufficiently high pH comprises a pH greater thanabout pH
 10. 120. The liquid formulation of claim 119, wherein saidsufficiently high pH comprises a pH greater than about pH
 11. 121. Theliquid formulation of claim 119, wherein said sufficiently high pHcomprises a pH greater than about pH 10, the formulation furthercharacterized in that the solution is suitable for lyophilization toform a lyophilate that can be reconstituted to form an aqueous solutionhaving a pH in the range about 6 to about 8 and the glibenclamideremains dissolved in solution.
 122. The liquid formulation of claim 119,wherein said sufficiently high pH comprises a pH greater than about pH11, the formulation further characterized in that the solution issuitable for lyophilization to form a lyophilate that can bereconstituted to form an aqueous solution having a pH in the range about6 to about 8 and the glibenclamide remains dissolved in solution. 123.The solid pharmaceutical composition of claim 104, wherein saidpharmaceutically inert compound is a bulking agent is selected from thegroup consisting of a mono-saccharide and di-saccharide.